EP1660436A1 - Inhibiteurs de la cathepsine - Google Patents

Inhibiteurs de la cathepsine

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Publication number
EP1660436A1
EP1660436A1 EP04761741A EP04761741A EP1660436A1 EP 1660436 A1 EP1660436 A1 EP 1660436A1 EP 04761741 A EP04761741 A EP 04761741A EP 04761741 A EP04761741 A EP 04761741A EP 1660436 A1 EP1660436 A1 EP 1660436A1
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EP
European Patent Office
Prior art keywords
cathepsin
compound
mixture
mammal
mmol
Prior art date
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EP04761741A
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German (de)
English (en)
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EP1660436A4 (fr
Inventor
Christopher Bayly
Cameron Black
Daniel J. Mckay
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Merck Canada Inc
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Merck Frosst Canada Ltd
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Publication of EP1660436A1 publication Critical patent/EP1660436A1/fr
Publication of EP1660436A4 publication Critical patent/EP1660436A4/fr
Withdrawn legal-status Critical Current

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    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/10—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by doubly bound oxygen or sulphur atoms
    • C07D295/104—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by doubly bound oxygen or sulphur atoms with the ring nitrogen atoms and the doubly bound oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
    • C07D295/108—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by doubly bound oxygen or sulphur atoms with the ring nitrogen atoms and the doubly bound oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain
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    • A61P19/00—Drugs for skeletal disorders
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    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C255/00—Carboxylic acid nitriles
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    • C07C255/24—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms containing cyano groups and singly-bound nitrogen atoms, not being further bound to other hetero atoms, bound to the same saturated acyclic carbon skeleton
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C255/00—Carboxylic acid nitriles
    • C07C255/01—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms
    • C07C255/24—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms containing cyano groups and singly-bound nitrogen atoms, not being further bound to other hetero atoms, bound to the same saturated acyclic carbon skeleton
    • C07C255/29—Carboxylic acid nitriles having cyano groups bound to acyclic carbon atoms containing cyano groups and singly-bound nitrogen atoms, not being further bound to other hetero atoms, bound to the same saturated acyclic carbon skeleton containing cyano groups and acylated amino groups bound to the carbon skeleton
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C255/00—Carboxylic acid nitriles
    • C07C255/45—Carboxylic acid nitriles having cyano groups bound to carbon atoms of rings other than six-membered aromatic rings
    • C07C255/46—Carboxylic acid nitriles having cyano groups bound to carbon atoms of rings other than six-membered aromatic rings to carbon atoms of non-condensed rings
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    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/23—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton
    • C07C323/39—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and nitrogen atoms, not being part of nitro or nitroso groups, bound to the same carbon skeleton at least one of the nitrogen atoms being part of any of the groups, X being a hetero atom, Y being any atom
    • C07C323/40—Y being a hydrogen or a carbon atom
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    • C07D223/02—Heterocyclic compounds containing seven-membered rings having one nitrogen atom as the only ring hetero atom not condensed with other rings
    • C07D223/06—Heterocyclic compounds containing seven-membered rings having one nitrogen atom as the only ring hetero atom not condensed with other rings with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/12—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms
    • C07D295/135—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms with the ring nitrogen atoms and the substituent nitrogen atoms separated by carbocyclic rings or by carbon chains interrupted by carbocyclic rings
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    • C07D307/26—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
    • C07D307/30—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D307/32—Oxygen atoms
    • C07D307/33—Oxygen atoms in position 2, the oxygen atom being in its keto or unsubstituted enol form
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    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
    • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
    • C07D333/06—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to the ring carbon atoms
    • C07D333/14—Radicals substituted by singly bound hetero atoms other than halogen
    • C07D333/20—Radicals substituted by singly bound hetero atoms other than halogen by nitrogen atoms
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    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
    • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
    • C07D333/06—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to the ring carbon atoms
    • C07D333/24—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06191—Dipeptides containing heteroatoms different from O, S, or N
    • C—CHEMISTRY; METALLURGY
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/02—Systems containing only non-condensed rings with a three-membered ring

Definitions

  • the invention relates generally to inhibitors of protein activity, and specifically to cathepsin inhibitors.
  • cathepsins belong to the papain superfamily of cysteine proteases. These proteases function in the normal physiological as well as pathological degradation of connective tissue. Cathepsins play a major role in intracellular protein degradation and turnover and remodeling. These cathepsins are naturally found in a wide variety of tissues. To date, a number of cathepsin have been identified and sequenced from a number of sources; for example, cathepsin B, F, H, L, K, S, W, and Z have been cloned.
  • Cathepsin K can be found in PCT Application WO 96/13523, Khepri Pharmaceuticals, Inc., published May 9, 1996, which is hereby incorporated by reference in its entirety.
  • Cathepsin L is implicated in normal lysosomal proteolysis as well as several diseases states, including, but not limited to, metastasis of melanomas.
  • Cathepsin S is implicated in Alzheimer's disease and certain autoimmune disorders, including, but not limited to juvenile onset diabetes, multiple sclerosis, pemphigus vulgaris, Graves' disease, myasthenia gravis, systemic lupus erythemotasus, rheumatoid arthritis and Hashimoto's thyroiditis; allergic disorders, including, but not limited to asthma; and allogenic immunbe responses, including, but not limited to, rejection of organ transplants or tissue grafts. Increased Cathepsin B levels and redistribution of the enzyme are found in tumors, suggesting a role in tumor invasion and matastasis.
  • Cathepsin B activity is implicated in such disease states as rheumatoid arthritis, osteoarthritis, pneumocystisis carinii, acute pancreatitis, inflammatory airway disease and bone and joint disorders.
  • Cysteine protease inhibitors such as E-64 (trans-epoxysuccinyl-L-leucylamide-(4- guanidino) butane) are known to be effective in inhibiting bone resorption. See Delaisse, J. M. et al., 1987, Bone 8:305-313, which is hereby incorporated by reference in its entirety.
  • the autosomal recessive disorder, pycnodysostosis characterized by an osteopetrotic phenotype with a decrease in bone resorption
  • pycnodysostosis characterized by an osteopetrotic phenotype with a decrease in bone resorption
  • all mutations identified in the cathepsin K gene are known to result in inactive protein. See Gelb, B. D. et al., 1996, Science 273:1236-1238; Johnson, M. R. et al, 1996, Genome Res 6:1050-1055, which are hereby incorporated by reference in their entirety. Therefore, it appears that cathepsin K is involved in osteoclast mediated bone resorption.
  • Cathepsin K is synthesized as a 37 kDa pre-pro enzyme, which is localized to the lysosomal compartment and where it is presumably autoactivated to the mature 27 kDa enzyme at low pH. See McQueney, M. S. et al, 1997, J Biol Chem 272:13955-13960; Littlewood-Evans, A. et al, 1997, Bone 20:81-86, which are hereby incorporated by reference in their entirety. Cathepsin K is most closely related to cathepsin S having 56 % sequence identity at the amino acid level.
  • the S 2 P 2 substrate specificity of cathepsin K is similar to that of cathepsin S with a preference in the PI and P2 positions for a positively charged residue such as arginine, and a hydrophobic residue such as phenylalanine or leucine, respectively. See Bromme, D. et al, 1996, J Biol Chem 271: 2126-2132; Bossard, M. J. et al., 1996, J Biol Chem 271:12517-12524, which are hereby incorporated by reference in their entirety.
  • Cathepsin K is active at a broad pH range with significant activity between pH 4-8, thus allowing for good catalytic activity in the resorption lacunae of osteoclasts where the pH is about 4-5.
  • Human type I collagen, the major collagen in bone is a good substrate for cathepsin K. See Kafienah, W., et al., 1998, Biochem J 331:727-732, which is hereby incorporated by reference in its entirety.
  • In vitro experiments using antisense oligonucleotides to cathepsin K have shown diminished bone resorption in vitro, which is probably due to a reduction in translation of cathepsin K mRNA.
  • inhibitors of Cathepsin K can reduce bone resorption.
  • Such inhibitors would be useful in treating disorders involving bone resorption, such as osteoporosis.
  • the invention provides compounds that are capable of treating and/or preventing cathepsin dependent conditions or disease states in a mammal in need thereof.
  • the invention provides compounds of the generic formula:
  • R is a non-hydrogen substituent with electron-withdrawing character adequate to, in conjunction with Ri, R 2 and R 3 , reduce the pKa of the nitrogen to ⁇ 6 in aqueous media
  • Ri is a substituent binding in the Si subsite of the active site of cathepsins
  • R 2 is a substituent binding in the S 2 subsite of the active site of cathepsins
  • R 3 is a substituent binding in the S 3 subsite of the active site of cathepsins.
  • Rx is a substituent binding in the Si subsite of the active site of cathepsins
  • R 2 is a substituent binding in the S 2 subsite of the active site of cathepsins
  • R 3 is a substituent binding in the S 3 subsite of the active site of cathepsins.
  • Residues that follow the scissile bond are called Pi', P 2 ', P 3 ⁇ etc. It has been found that the main chain of protein inhibitors having very different overall structure are highly similar in the region between P 3 and P 3 ' with especially high similarity for P 2 , Pi and Pi'. It is generally accepted that each protease active site has subsites Si, S 2 , etc. that receive the side groups of residues Pi, P 2 , etc. of the substrate or inhibitor and subsites Si', S 2 ', etc. that receive the side groups of Pi', P 2 ', etc. of the substrate or inhibitor.
  • This fragment is thus capable of making an important neutral hydrogen bond to a cathepsin glycine conserved amongst the entire papain family of cathepsins.
  • Both amides and anilines are capable of making hydrogen bonds to carbonyl groups, but have metabolic liabilities.
  • the ethylamine is basic and is protonated in biological systems, generating a charge that reduces binding to the target enzyme. With sufficiently electron-withdrawing R (or more specifically trifluoroethylamine), the amine is not protonated and provides a superior hydrogen bond to acceptor oxygen in the cathepsin active site.
  • cathepsins an important acceptor oxygen is situated in between the S 2 and S 3 subsites of the active site; in cathepsin K this is Gly66.
  • This residue is a conserved residue amongst this entire papain family of proteins, including cathepsins B, F, H, K, L, L2, O, S, W, and Z, falcipain, falcipain-1 and falcipain 2.
  • using the non-basic ethylamine linker to join inhibitor fragments binding the S 2 and S 3 subsites of cathepsins allows for more potent inhibition than the corresponding amides.
  • nonhydrogen atoms each independently selected from C, O, N, S, P, F, Cl,
  • R is a non-hydrogen electron-withdrawing substituent such that, together with R l5 R 2 and R 3 , the basicity of the nitrogen is lowered to less than a pKa of 6; wherein a molecule of the composition is interacting with a cathepsin such that the CH-NH region in the chemical formula is interacting favorably with the cathepsin between S 2 and S 3 , Ri interacts favorably with Si but not S 3 of the cathepsin active site, R 2 interacts favorably with S 2 but not S 3 of the cathepsin active site, and R 3 interacts favorably with S 3 but not S 2 or Si of the cathepsin active site.
  • the cathepsin is selected from cathepsin B, F, H, K, L, L2, O, S, W and Z.
  • the cathepsin is selected from cathepsin K, L, S, and B.
  • the cathepsin is cathepsin L.
  • the cathepsin is cathepsin S.
  • the cathepsin is cathepsin B.
  • the cathepsin is cathepsin F.
  • R 4 does not interact favorably with subsites S 2 , S 3 and Si, respectively, of a cathepsin active site.
  • R is selected from -CF 3 , -CHF , -CH 2 F, -CF 2 R 5 , and -CHF R s , wherein R 5 is C ⁇ -6 alkyl, aryl, or heteroaryl optionally substituted with 1 to 4 substituents selected from halo, C ⁇ -3 alkyl, . 3 alkoxy, hydroxy, hydroxyalkyl, keto, cyano, heterocyclyl, C 3 .
  • R 2 has at least one carbon or sulfur atom which simultaneously fulfills the following three distance critieria: it is within 7 A of C ⁇ 2 ⁇ , and it is within 8.5 A of C ⁇ 8 and it is within 7 A of C ⁇ 34 of a cathepsin.
  • R 2 comprises nonpolar regions, i another class of the invention, R 2 comprises lipophilic regions.
  • Ri comprises a region that stably fits into subsite Si of a cathepsin active site, having at least one carbon atom within 5 A of C01 25 of a cathepsin.
  • Ri is non-immunogenic
  • R 3 has at least one carbon or sulfur atom which simultaneously fulfills the following two distance critieria: it is within 5.5 A of C ⁇ 66 , and it is within 7 A of C ⁇ 6 o of a cathepsin.
  • R 3 comprises nonpolar regions
  • hi another class of the invention R 3 comprises lipophilic regions.
  • the nitrogen has a pKa of less than 6 and makes a hydrogen bond with the cathepsin amide carbonyl of glycine 66 of a cathepsin.
  • the compound has a molecular weight of less than 1000 daltons. Ii a class of the invention, the compound forms a covalent bond with cysteine 25 of a cathepsin. h a class of the invention, the compound binds to the active site of a cathepsin with an IC50 of less than 10 micromolar in a purified enzyme assay.
  • the pKa of the nitrogen of the secondary amine shown in claim 1 is ⁇ 5 in an aqueous medium.
  • lipophilic refers to a compound, which, as a separate entity, is more soluble in nonpolar solvents (e.g. cyclohexane) than water.
  • lipophilic group in the context of being attached to a molecule, refers to a region of the molecule having high hydrocarbon content thereby giving the group high affinity to nonpolar solvents or lipid phases.
  • a lipophilic group can be, for example, an alkyl or cycloalkyl chain(preferably n-alkyl) of less than 30 carbons.
  • lipophilic groups include the alkyl chains attached to naturally-occurring and synthetic aromatic and non-aromatic moieties such as fatty acids, esters and alcohols, and other lipid molecules.
  • Other examples of lipophilic molecules are cage structures such as adamantane and buckminsterfullerenes, and aromatic hydrocarbons such as benzene, perylene, phenanthrene, anthracene, naphthalene, pyrene, chrysene, and naphthacene.
  • lipophilic group as used herein are the carbons and attached hydrogens of alkyl chains, cycloalkyl chains, aryl rings, or heteroaryl rings.
  • lipophilic group as used herein also includes divalent sulfur.
  • substantially homologous when used in connection with amino acid sequences, refers to sequences which are substantially identical to or similar in sequence, giving rise to a homology in conformation and thus to similar biological activity. The term is not intended to imply a common evolution of the sequences.
  • substantially homologous sequences are at least 50% more preferably at least 80% identical in sequence, at least over any regions known to be involved in the desired activity. Most preferably, no more than five residues, other than at the termini, are different.
  • the divergence in sequence, at least in the aforementioned regions is in the form of "conservative modifications".
  • Constant modifications are defined as (a) conservative substitutions of amino acids as hereafter defined; and (b) single or multiple insertions or deletions of amino acids at the termini, at interdomain boundaries, in loops or in other segments of relatively high mobility (as indicated, e.g., by the failure to clearly resolve their structure upon X-ray diffraction analysis or NMR).
  • conservative substitutions of amino acids as hereafter defined
  • single or multiple insertions or deletions of amino acids at the termini, at interdomain boundaries, in loops or in other segments of relatively high mobility as indicated, e.g., by the failure to clearly resolve their structure upon X-ray diffraction analysis or NMR.
  • no more than about five amino acids are inserted or deleted at a particular locus, and the modifications are outside regions known to contain binding sites important to activity.
  • Conservative substitutions are herein defined as exchanges within one of the following five groups: (1) small aliphatic, nonpolar or slightly polar residues: Ala, Ser, Thr (Pro, Gly); (2) polar, negatively charged residues: and their amides Asp, Asn, Glu, Gin; (3) polar, positively charged residues: His, Arg, Lys; (4) large, aliphatic, nonpolar residues: Met, Leu, De, Val (Cys); and (5) large, aromatic residues: Phe, Tyr, Tip.
  • Residues Pro, Gly and Cys are parenthesized because they have special conformational roles. Cys participates in formation of disulfide bonds. Gly imparts flexibility to the chain.
  • Pro imparts rigidity to the chain and disrupts alpha helices. These residues may be essential in certain regions of the polypeptide, but substitutable elsewhere.
  • Semi-conservative substitutions are defined to be exchanges between two of groups (l)-(5) above which are limited to supergroup (a), comprising (1), (2) and (3) above, or to supergroup (b), comprising (4) and (5) above.
  • the term "cathepsin” or “cathepsins” as used herein refers to enzymes belonging to the papain family, i.e. where the active form of the enzyme is folded similarly to papain ( conserveed Domain smart00645.7 in the NCBI conserveed Domain Database http:
  • cathepsins in human, mouse, rabbit, primates, rat, and plasmodiumfalciparum are being referred to herein.
  • the human cathepsins specifically included are B, F, H, K, L, L2, O, S, W, and Z; also specifically included are enzymes in mouse, rabbit, primates, and rat exhibiting greater than 80% sequence identity (comparing the active forms of the enzymes) to the most similar of the preceding human cathepsins.
  • falcipain, falcipain-1 and falcipain-2 from plasmodiumfalciparum and any other enzymes from plasmodiumfalciparum exhibiting greater than 80% sequence identity to the most similar of these, again comparing the active forms of the enzymes.
  • a specific amino acid residue(e.g. glycine 66) or a C ⁇ (e.g. C ⁇ 66 ) in a cathepsin is referred to herein via a combination of the residue numbering used for the active form of Cathepsin K and the primary sequence alignment given in Figure 1. This sequence alignment is for the relevant portion of the active form of the protein for human cathepsins and falcipains as labeled in the Figure.
  • residue numbers referred to herein are given, formatted vertically above the aligned sequences. Other residue numbers referred to can found by counting along from the closest numbered residue or by direct reference to the Cathepsin K sequence. The numbering is obtained based on the SWISSPROT primary accession #P43235 for human cathepsin K, where the first residue in the CHAIN, the active form of the protein (residue number 115 in #P43235) is reset herein to be residue number 1.
  • sequence alignment herein is used to generalize the reference to a Cathepsin K residue to the other human cathepsins and the falcipains.
  • residues in mouse, rabbit, primates, and rat cathepsins is implied by a standard primary sequence alignment, using default parameters, to the most highly homologous sequence specified below: 22 56
  • CathepsinK YPYVG QEESCMYNPT GKAAKCR
  • CathepsinB YESFTVGCRPYSIPPCEHHVNGSRPPCTGEGDTPKCSKICEPGYSPTYKQD
  • CathepsinF YSYQG HMQSCNFSAE KAKVYIN
  • CathepsinH YPYQG KDGYCKFQPG KAIGFVK
  • CathepsinL YPYEA TEESCKYNPK YSVANDT
  • CathepsinS YPYKA -MDQKCQYDSK YRAATCS
  • CathepsinW YPFQGK- VRAHRCHPKKY QKVAWIQ
  • CathepsinZ NNYQAKDQECDKFNQCGTCNEFKE CHAIRNY
  • CathepsinL2 YPYVA VDEICKYRPE NSVANDT
  • CathepsinK GYREIPEG — NEKALKRAVARVGPVSV0EDASLTSFQFYSKGVYYDES
  • CathepsinB KHYGYNSYSVSNSEKDIMAEIYKNGPVEG ⁇ SVYSD-FLLYKSGVYQFTVT
  • CathepsinF DSVELSQ NEQKLAAWLAKRGPISV ⁇ NAFGMQFYRHGISRPLRPL
  • CathepsinH DVANTTIY— -DEEAMVEAVALYNPVS ⁇ FEVTQD-FMMYRTGIYSSTS
  • CathepsinL GFVDIPK QEKALMKAVATVGPISVEpDAGHESFLFYKEGIYFEPD
  • CathepsinO KGYSAYDFSD--QEDEMAKAI TFGPLVVJJVDAVSWQDYLGGHQHHCSS
  • CathepsinS KYTELPYG — REDVLKEAVANKGPVSVgjVDARHPSFFLYRSGVYYEPS
  • CathepsinW DFIMLQN NEHRIAQYLATYGPJ V01NMKPLQLYRKGVIKATPTT
  • CathepsinZ TLWRVGDYGSLSGREKMMAEIYANGPISC ⁇ ATER-LANYTGGIYAEYQ
  • CathepsinL2 GFTVVAPG — KEKALMKAVATVGPISV0MDAGHSSFQFYKSGIYFEPD
  • Falcipain3 NYLSVPD -N-KLKEALRFLGPISJ VAVSDD-FAFYKEGIFDGEC CathepsinK : CNS — -DNLNHAVLAVGYGIQKG
  • CathepsinB GEMMG GHAIRILGWGVENG
  • CathepsinF CSP — -WLIDHAVLLVGYGNRSD
  • CathepsinH CHKTP— DKVNHAVLAVGYGEKNG
  • CathepsinO GE ANHAVLLTGFDKTGS
  • CathepsinW CDP QLVDHSVI VGFGSVKSEEGIWAETVSS QSQPQPPHP
  • CathepsinZ DTT YINHVVSVAGWGISDG
  • CathepsinK NKHWIIKNSWGENWGNKGY MARNKN NACGIANLASFPKM-
  • CathepsinB TPYWLVANSWNTDWGDNGFFKILRGQD HCGIESEVVAGIPRT
  • CathepsinF VPFWAIKNSWGTDWGEKGYYYLHRGSG ACGVNTMASSAVVD-
  • CathepsinL NKYWLVKNSWGEEWGMGGYVKMAKDRR NHCGIASAASYPTV-
  • CathepsinO TPYWIVRNSWGSSWGVDGYAHVKMGSN VCGIADSVSSIFV--
  • CathepsinS KEYWLVKNSWGHNFGEEGYIRMARNKG NHCGIASFPSYPEI-
  • CathepsinW TPYWILKNSWGAQWGEKGYFRLHRGSN -TCGLTKFPLTARVQK
  • CathepsinZ TEYWIVRNSWGEPWGERGWLRIVTSTYKDGKGARYNLAIEEHCTFGDPIV
  • CathepsinL2 SKYWLVKNSWGPEWGSNGYVKIAKDKN NHCGIATAASYPNV--
  • Falcipain IYYWIIKNSWSKKWGENGFMRLSRNKNGD — NVFCGIGEEVFYPIL-- Falcipain2 : FYYYIIKNSWGSDWGEGGYINLETDENGY — KKTCSIGTEAYVPLLE- Falcipain3 : HYYYUKNSWGQQWGERGFINIETDESGL— MRKCGLGTD AFIPLIE- CathepsinK : (SEQ ID NO: 1)
  • CathepsinB D (SEQ ID NO: 2)
  • CathepsinF (SEQ ID NO: 3)
  • CathepsinH (SEQ ID NO: 4)
  • CathepsinL (SEQ ID NO: 5)
  • CathepsinO (SEQ ID NO: 6)
  • CathepsinS (SEQ ID NO: 7)
  • CathepsinW PDMKPRVSCPP-(SEQ ID NO: 8)
  • CathepsinZ (SEQ ID NO: 9)
  • CathepsinL2 (SEQ ID NO: 10)
  • vorably refers to the favorable result of a molecular modeling calculation in which a computer model of the ligand molecule is placed into the active site of a computer model of the cathepsin, making a ligand:cathepsin complex, and the ligand: cathepsin complex is energy minimized using a standard molecular mechanics forcefield such as MMFF94s (T. A. Halgren, Journal of Computational Chemistry (1999), 20, pp. 720-729.).
  • the initial placement of the ligand into the active site is done so as to optimally allow the ligand:enzyme interactions between the substituents of the ligand and the subsites of the enzyme as stated herein.
  • the computer model of the active site is based on an Xray crystal structure (e.g. Protein Databank entry 1MEM for cathepsin K) if available for the cathepsin of interest (minus the non-protein atoms), or if an Xray crystal structure is not available, one can use a homology-built structure based on the most similar cathepsin for which an Xray structure is available, or simply use directly the most similar cathepsin for which an Xray structure is available.
  • an Xray crystal structure e.g. Protein Databank entry 1MEM for cathepsin K
  • a "favorable result of a modeling calculation” means that after the energy minimization is complete there are no bad steric interactions between the ligand and the active site and there are energy-stabilising hydrogen bonding and lipophilic interactions between the ligand and the active site. If it is believed that the ligand forms a covalent bond with the active site sulfur of cysteine-25, the energy minimization may include that covalent bond in the modeled ligandxathepsin complex submitted to the calculation. Interactions between the ligand and the cathepsin as claimed herein are based on the geometry of the ligand: cathepsin complex that would result from such an energy minimization.
  • favorable interactions between a substituent of the ligand and S 2 require having at least one carbon or divalent sulfur atom of the substituent simultaneously fulfilling the following three distances: it is within 7 A of C ⁇ 26 , and it is within 8.5 A of C ⁇ 68 and it is within 7 A of C ⁇ i 3 of the cathepsin.
  • favorable interactions between a substituent of the ligand and S 2 require having at least one atom of a lipophilic group of the substituent within 5.5 A of atoms of two of residues 67, 68, 134.
  • favorable interactions between a substituent of the ligand and S 2 require having hydrogen bonding between the substituent and glutamate 209 of cathepsin B.
  • favorable interactions between a substituent of the ligand and S 3 require having at least one carbon or divalent sulfur atom of the substituent simultaneously fulfilling the following two distances: it is within 5.5 A of C 66 , and it is within 7 A of C ⁇ 6 o of the cathepsin.
  • favorable interactions between a substituent of the ligand and S 3 require having at least one atom of a lipophilic group of the substituent within 5.5 A of Cage and atoms of either residue 60 or 61. In another embodiment of the invention, favorable interactions between a substituent of the ligand and S 3 require having at least one atom of a lipophilic group of the substituent within 5.5 A of either residue 60 or 61.
  • favorable interactions between a substituent of the ligand and S x require having at least one carbon atom of the substituent within 5 A of C ⁇ 25 - hi another embodiment of the invention, favorable interactions between a substituent of the ligand and Si require having a covalent bond between the active site cysteine sulfur of residue 25 and an electrophilic carbon of the ligand, preferably a carbonyl or nitrile carbon.
  • favorable interactions between the CH-NH region in the chemical formula of the ligand and the cathepsin between S2 and S3 requires having the N of the chemical formula within 4 A of the peptide oxygen of Gly66, and the C of the chemical formula (referring only the C directly connected with R ) within 5.5 A of C ⁇ 66 .
  • favorable interactions between the CH-NH region in the chemical formula of the ligand and the cathepsin between S2 and S3 requires having the NH of the chemical formula hydrogen bonding to the amide O of residue 66.
  • alkyl is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having one to ten carbon atoms unless otherwise specified.
  • Cl-Cio as in “C1-C10 alkyl” is defined to include groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbons in a linear, branched, or cyclic arrangement.
  • C ⁇ -C ⁇ o alkyl specifically includes methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, and so on.
  • alkyloxy represents an alkyl group as defined above, unless otherwise indicated, wherein said alkyl group is attached through an oxygen bridge.
  • cycloalkyl or “carbocycle” shall mean cyclic rings of alkanes of three to eight total carbon atoms, unless otherwise indicated, or any number within this range (i.e., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl).
  • aryl is intended to mean any stable monocyclic or bicyclic carbon ring of up to 12 atoms in each ring, wherein at least one ring is aromatic.
  • aryl elements examples include phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthryl, anthryl or acenaphthyl.
  • aryl substituent is bicyclic and one ring is non-aromatic, it is understood that attachment is via the aromatic ring.
  • heteroaryl represents a stable monocyclic, bicyclic or tricyclic ring of up to 10 atoms in each ring, wherein at least one ring is aromatic and contains from 1 to 4 heteroatoms selected from the group consisting of O, N and S.
  • Heteroaryl groups within the scope of this definition include but are not limited to: benzoimidazolyl, benzofuranyl, benzofurazanyl, benzopyrazolyl, benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazolyl, carbolinyl, cinnolinyl, furanyl, indolinyl, indolyl, indolazinyl, indazolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthpyridinyl, oxadiazolyl, oxazolyl, oxazoline, isoxazoline, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridopyridinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazoliny
  • heteroaryl substituent is bicyclic and one ring is non-aromatic or contains no heteroatoms, it is understood that attachment is via the aromatic ring or via the heteroatom containing ring, respectively. If the heteroaryl contains nitrogen atoms, it is understood that the corresponding N-oxides thereof are also encompassed by this definition.
  • halo or “halogen” as used herein is intended to include chloro, fluoro, bromo and iodo.
  • alkoxy as used herein means an alkyl portion, where alkyl is as defined above, connected to the remainder of the molecule via an oxygen atom.
  • alkoxy examples include methoxy, ethoxy and the like.
  • hydroxyalkyl means a linear monovalent hydrocarbon raidcal of one to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbons substituted with one or two hydroxy groups, provided that if two hydroxy groups are present they are not both on the same carbon atom. Representative examples include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 2- hydroxypropyl, 3- hydroxypropyl, and the like.
  • heterocycle or “heterocyclyl” as used herein is intended to mean a 5- to 10- membered nonaromatic ring, unless otherwise specified, containing from 1 to 4 heteroatoms selected from the group consisting of O, N, S, SO, or S0 2 and includes bicyclic groups.
  • Heterocyclyl therefore includes, but is not limited to the following: piperazinyl, piperidinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropiperidinyl, tetrahydrothiophenyl and the like.
  • the present invention also includes N-oxide derivatives and protected derivatives of compounds of Formula I.
  • compounds of Formula I when compounds of Formula I contain an oxidizable nitrogen atom, the nitrogen atom can beconverted to an N-oxide by methods well known in the art.
  • compounds of Formula I when compounds of Formula I contain groups such as hydroxy, carboxy, tbiol or anygroup containing a nitrogen atom(s), these groups can be protected with a suitable protecting groups.
  • a comprehensive list of suitable protective groups can be found in T.W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, Inc.
  • the protected derivatives of compounds of Formula I can be prepared by methods well known in the art. . Also included within the scope of the present invention is a pharmaceutical composition which is comprised of a compound as described above and a pharmaceutically acceptable carrier. The invention is also contemplated to encompass a pharmaceutical composition, which is comprised of a pharmaceutically acceptable carrier and any of the compounds specifically disclosed in the present application. These and other aspects of the invention will be apparent from the teachings contained herein.
  • the pharmaceutically acceptable salts of the compounds of this invention include the conventional non-toxic salts of the compounds of this invention as formed inorganic or organic acids.
  • non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like, as well as salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic and the like.
  • inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like
  • organic acids such as acetic, propionic, succinic, glycolic, ste
  • the preparation of the pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts is more fully described by Berg et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977:66:1- 19, hereby incorporated by reference.
  • the pharmaceutically acceptable salts of the compounds of this invention can be synthesized from the compounds of this invention which contain a basic or acidic moiety by conventional chemical methods. Generally, the salts of the basic compounds are prepared either by ion exchange chromatography or by reacting the free base with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid in a suitable solvent or various combinations of solvents. Similarly, the salts of the acidic compounds are formed by reactions with the appropriate inorganic or organic base.
  • the compounds of the present invention are inhibitors of cathepsins and are therefore useful to treat or prevent cathepsin dependent diseases or conditions in mammals, preferably humans.
  • the compounds of the present invention are inhibitors of Cathepsin K and are therefore useful to treat or prevent Cathepsin K dependent diseases or conditions in mammals, preferably humans.
  • Cathepsin dependent diseases or conditions refers to pathologic conditions that depend on the activity of one or more cathepsins.
  • Cathepsin K dependent diseases or conditions refers to pathologic conditions that depend on the activity of Cathepsin K.
  • Diseases associated with Cathepsin K activities include osteoporosis, glucocorticoid induced osteoporosis, Paget's disease, abnormally increased bone turnover, periodontal disease, tooth loss, bone fractures, rheumatoid arthritis, osteoarthritis, periprosthetic osteolysis, osteogenesis imperfecta, obesity, atherosclerosis, chronic obstructive pulmonary disease, juvenile onset diabetes, multiple sclerosis, pemphigus vulgaris, Graves' disease, myasthenia gravis, systemic lupus erythemotasus, rheumatoid arthritis and Hashimoto's thyroiditis, asthma, allogenic immune responses, parasitic infection, cancer, metastatic bone disease, hypercalcemia of malignancy or multiple myeloma.
  • An embodiment of the invention is a method of inhibiting cathepsin activity in a mammal in need thereof, comprising administering to the mammal a therapeuticaily effective amount of any of the compounds or any of the pharmaceutical compositions described above.
  • a class of the embodiment is the method wherein the cathepsin activity is cathepsin K activity.
  • Another embodiment of the invention is a method of treating or preventing cathepsin dependent conditions in a mammal in need thereof, comprising administering to the mammal a therapeuticaily effective amount of any of the compounds or any of the pharmaceutical compositions described above.
  • a class of the embodiment is the method wherein the cathepsin activity is cathepsin K activity.
  • Another embodiment of the invention is a method of inhibiting bone loss in a mammal in need thereof, comprising administering to the mammal a therapeuticaily effective amount of any of the compounds or any of the pharmaceutical compositions described above.
  • Another embodiment of the invention is a method of reducing bone loss in a mammal in need thereof, comprising administering to the mammal a therapeuticaily effective amount of any of the compounds or any of the pharmaceutical compositions described above.
  • the utility of cathepsin K inhibitors in the inhibition of bone resorption is known in the literature. See Stroup G.B. et al., "Potent and selective inhibition of human cathepsin K leads to inhibition of bone resorption in vivo in a nonhuman primate.” J. Bone Miner. Res., 16:1739- 1746;2001; and Votta, BJ.
  • Another embodiment of the invention is a method of treating or preventing osteoporosis in a mammal in need thereof, comprising administering to the mammal a therapeuticaily effective amount of any of the compounds or any of the above pharmaceutical compositions described above.
  • the utility of cathepsin K inhibitors in the treatment or prevention of osteoporosis is known in the literature. See, Saftig P. et al, "Impaired osteoclast bone resorption leads to osteoporosis in cathepsin K-deficient mice.” Proc.
  • Another embodiment of the invention is a method of treating or preventing rheumatoid arthritic condition in a mammal in need thereof, comprising administering to the mammal a therapeuticaily effective amount of any of the compounds or any of the pharmaceutical compositions described above.
  • RA rheumatoid arthritis
  • cathepsin K positive osteoclasts are the cell types that mediate the focal bone resorption associated with rheumatoid synovial lesion. See Hou, W-S et al., "Comparision of Cathepsin K and S expression within the Rheumatoid and Osteoarthritic Synovium", Arthritis Rheumatism 46: 663-74, 2002.
  • generalized bone loss is a major cause of morbility associated with severe RA.
  • the frequency of hip and spinal fractures is substantially increased in patients with chronic RA.
  • Another embodiment of the invention is a method of treating or preventing the progression of osteoarthritis in a mammal in need thereof, comprising administering to the mammal a therapeuticaily effective amount of any of the compounds or any of the pharmaceutical compositions described above.
  • osteoarthritis is accompanied with a well-defined changes in the joints, including erosion of the articular cartilage surface, peri-articular endochondral ossification/osteophytosis, and subchondral bony sclerosis and cyst formation, see Oettmeier R & Abendroth, K, " Osteoarthritis and bone: osteologic types of osteoarthritis of the hip", Skeletal Radiol. 18: 165-74, 1989. Recently, the potential contribution of subchondral bone sclerosis to the initiation and progression of OA have been suggested.
  • Stiffened subchondral bone as the joint responding to repetitive impulsive loading is less able to attenuate and distribute forces through the joint, subjecting it to greater mechanical stress across the articular cartilage surface. This in turn accelerates cartilage wear and fibrillate. See, Radin, EL and Rose RM, "Role of subchondral bone in the initiation and progression of cartilage damage.” Clin. Orthop. 213: 34-40, 1986. Inhibition of excessive subarticular bone resorption by an anti-resorptive agent such as a cathepsin K inhibitor, will lead to inhibition of subchondral bone turnover, thus may have a favorable impact on OA progression.
  • an anti-resorptive agent such as a cathepsin K inhibitor
  • cathepsin K protein expression was recently identified in synovial fibroblasts, macrophage-like cells, and chondrocytes from synovium and articular cartilage specimens derived from OA patients. See, Hou, W-S et al., "Comparison of Cathepsin K and S expression within the Rheumatoid and Osteoarthritic Synovium", Arthritis Rheumatism 46: 663-74, 2002; and Dodd RA et al, "Expression of Cathepsin K messenger RNA in giant cells and their precursors in human osteoarthritic synovial tissues".
  • cathepsin K inhibitors in the treatment or prevention of osteoarthritis as described in this invention thus comprise of two different mechanisms, one is on the inhibition of osteoclast-driven subchondral bone turnover, and two is on the direct inhibition of collagen type II degeneration in the synovium and cartilage of patients with OA.
  • Another embodiment of the invention is a method treating cancer in a mammal in need thereof, comprising administering to the mammal a therapeuticaily effective amount of any of the compounds or any of the pharmaceutical compositions described above. It is known in the literature that cathepsin K is expressed in human breast carcinoma.
  • Exemplifying the invention is the use of any of the compounds described above in the preparation of a medicament for the treatment and/or prevention of osteoporosis in a mammal in need thereof. Still further exemplifying the invention is the use of any of the compounds described above in the preparation of a medicament for the treatment and/or prevention of: bone loss, bone resorption, bone fractures, metastatic bone disease and/or disorders related to cathepsin functioning.
  • the compounds of this invention may be administered to mammals, preferably humans, either alone or, preferably, in combination with pharmaceutically acceptable carriers or diluents, optionally with known adjuvants, such as alum, in a pharmaceutical composition, according to standard pharmaceutical practice.
  • the compounds can be administered orally or parenterally, including the intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical routes of administration.
  • carriers which are commonly used include lactose and corn starch, and lubricating agents, such as magnesium stearate, are commonly added.
  • useful diluents include lactose and dried corn starch.
  • the selected compound may be administered, for example, in the form of tablets or capsules, or as an aqueous solution or suspension.
  • the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like;
  • the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like.
  • suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like.
  • Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.
  • Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
  • the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring agents may be added.
  • emulsifying and suspending agents emulsifying and suspending agents.
  • certain sweetening and/or flavoring agents may be added.
  • sterile solutions of the active ingredient are usually prepared, and the pH of the solutions should be suitably adjusted and buffered.
  • the total concentration of solutes should be controlled in order to render the preparation isotonic.
  • the compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles.
  • Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
  • Compounds of the invention may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled.
  • the compounds of the present invention may also be coupled with soluble polymers as targetable drug carriers.
  • Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxy-ethylaspartamide-phenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues.
  • the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymers of polyactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked or amphipathic block copolymers of hydrogels.
  • a drug for example, polylactic acid, polyglycolic acid, copolymers of polyactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked or amphipathic block copolymers of hydrogels.
  • the instant compounds are also useful in combination with known agents useful for treating or preventing osteoporosis, glucocorticoid induced osteoporosis, Paget's disease, abnormally increased bone turnover, periodontal disease, tooth loss, bone fractures, rheumatoid arthritis, osteoarthritis, periprosthetic osteolysis, osteogenesis imperfecta, obesity, atherosclerosis, chronic obstructive pulmonary disease, juvenile onset diabetes, multiple sclerosis, pemphigus vulgaris, Graves' disease, myasthenia gravis, systemic lupus erythemotasus, rheumatoid arthritis and Hashimoto's thyroiditis, asthma, allogenic immune responses, parasitic infection, cancer, metastatic bone disease, hypercalcemia of malignancy or multiple myeloma.
  • Combinations of the presently disclosed compounds with other agents useful in treating or preventing osteoporosis or other bone disorders are within the scope of the invention.
  • agents include the following: an organic bisphosphonate; an estrogen receptor modulator; an androgen receptor modulator; an inhibitor of osteoclast proton ATPase; an inhibitor of HMG-CoA reductase; an integrin receptor antagonist; an osteoblast anabolic agent, such as PTH; and the pharmaceutically acceptable salts and mixtures thereof.
  • a preferred combination is a compound of the present invention and an organic bisphosphonate.
  • Another preferred combination is a compound of the present invention and an estrogen receptor modulator. Another preferred combination is a compound of the present invention and an androgen receptor modulator. Another preferred combination is a compound of the present invention and an osteoblast anabolic agent.
  • Organic bisphosphonate includes, but is not limited to, compounds of the chemical formula
  • n is an integer from 0 to 7 and wherein A and X are independently selected from the group consisting of H, OH, halogen, NH2, SH, phenyl, C1-C30 alkyl, C3-C30 branched or cycloalkyl, bicyclic ring structure containing two or three N, C1-C30 substituted alkyl, C1-C10 alkyl substituted NH2, C3- C10 branched or cycloalkyl substituted NH2, C1-C10 dialkyl substituted NH2, C1-C10 alkoxy, C1-C10 alkyl substituted tbio, thiophenyl, halophenylthio, C1-C10 alkyl substituted phenyl, pyridyl, furanyl, pyrrolidinyl, imidazolyl, imidazopyridinyl, and benzyl, such that both A and X are not selected from H or OH when n is
  • the alkyl groups can be straight, branched, or cyclic, provided sufficient atoms are selected for the chemical formula.
  • the C1-C30 substituted alkyl can include a wide variety of substituents, nonlimiting examples which include those selected from the group consisting of phenyl, pyridyl, furanyl, pyrrolidinyl, imidazonyl, NH2, C1-C10 alkyl or dialkyl substituted NH2, OH, SH, and C1-C10 alkoxy.
  • the foregoing chemical formula is also intended to encompass complex carbocyclic, aromatic and hetero atom structures for the A and/or X substituents, nonlimiting examples of which include naphthyl, quinolyl, isoquinolyl, adamantyl, and chlorophenylthio.
  • Pharmaceutically acceptable salts and derivatives of the bisphosphonates are also useful herein.
  • Non-limiting examples of salts include those selected from the group consisting alkali metal, alkaline metal, ammonium, and mono-, di-, tri-, or tetra-Cl-C30-alkyl-substituted ammonium.
  • Preferred salts are those selected from the group consisting of sodium, potassium, calcium, magnesium, and ammonium salts.
  • Non-limiting examples of derivatives include those selected from the group consisting of esters, hydrates, and amides. It should be noted that the terms "bisphosphonate” and “bisphosphonates”, as used herein in referring to the therapeutic agents of the present invention are meant to also encompass diphosphonates, biphosphonic acids, and diphosphonic acids, as well as salts and derivatives of these materials. The use of a specific nomenclature in referring to the bisphosphonate or bisphosphonates is not meant to limit the scope of the present invention, unless specifically indicated.
  • a specific weight or percentage of a bisphosphonate compound in the present invention is on an acid active weight basis, unless indicated otherwise herein.
  • the phrase "about 5 mg of a bone resorption inhibiting bisphosphonate selected from the group consisting of alendronate, pharmaceutically acceptable salts thereof, and mixtures thereof, on an alendronic acid active weight basis” means that the amount of the bisphosphonate compound selected is calculated based on 5 mg of alendronic acid.
  • Non-limiting examples of bisphosphonates useful herein include the following: Alendronate, also known as alendronic acid, alendronate sodium, alendronate monosodium trihydrate or 4-amino-l-hydroxybutylidene-l,l-bisphosphonic acid monosodium trihydrate.
  • Alendronate is described in U.S. Patents 4,922,007, to Kieczykowski et al., issued May 1, 1990; 5,019,651, to Kieczykowski et al., issued May 28, 1991; 5,510,517, to Dauer et al., issued April 23, 1996; 5,648,491, to Treasure et al., issued July 15, 1997, all of which are incorporated by reference herein in their entirety.
  • 1,1-dichloromethylene- 1,1 -diphosphonic acid (clodronic acid), and the disodium salt (clodronate, Procter and Gamble), are described in Belgium Patent 672,205 (1966) and J. Org. Chem 32, 4111 (1967), both of which are incorporated by reference herein in their entirety.
  • Nonlimiting examples of bisphosphonates include alendronate, cimadronate, clodronate, etidronate, ibandronate, incadronate, minodronate, neridronate, olpadronate, pamidronate, piridronate, risedronate, tiludronate, and zolendronate, and pharmaceutically acceptable salts and esters thereof.
  • a particularly preferred bisphosphonate is alendronate, especially a sodium, potassium, calcium, magnesium or ammonium salt of alendronic acid. Exemplifying the preferred bisphosphonate is a sodium salt of alendronic acid, especially a hydrated sodium salt of alendronic acid.
  • the salt can be hydrated with a whole number of moles of water or non whole numbers of moles of water. Further exemplifying the preferred bisphosphonate is a hydrated sodium salt of alendronic acid, especially when the hydrated salt is alendronate monosodium trihydrate. It is recognized that mixtures of two or more of the bisphosphonate actives can be utilized.
  • the precise dosage of the organic bisphosphonate will vary with the dosing schedule, the particular bisphosphonate chosen, the age, size, sex and condition of the mammal or human, the nature and severity of the disorder to be treated, and other relevant medical and physical factors. Thus, a precise pharmaceutically effective amount cannot be specified in advance and can be readily determined by the caregiver or clinician.
  • an appropriate amount of bisphosphonate is chosen to obtain a bone resorption inhibiting effect, i.e. a bone resorption inhibiting amount of the bisphosphonate is administered.
  • an effective oral dose of bisphosphonate is typically from about 1.5 to about 6000 ⁇ g/kg body weight and preferably about 10 to about 2000 ⁇ g/kg of body weight.
  • common human doses which are administered are generally in the range of about 2 mg/day to about 40 mg/day, preferably about 5 mg/day to about 40 mg/day. In the U.S.
  • presently approved dosages for alendronate monosodium trihydrate are 5 mg/day for preventing osteoporosis, 10 mg/day for treating osteoporosis, and 40 mg/day for treating Paget's disease.
  • the bisphosphonate can be administered at intervals other than daily, for example once-weekly dosing, twice-weekly dosing, biweekly dosing, and twice-monthly dosing.
  • alendronate monosodium trihydrate would be administered at dosages of 35 mg/week or 70 mg/week.
  • Selective estrogen receptor modulators refers to compounds which interfere or inhibit the binding of estrogen to the receptor, regardless of mechanism.
  • estrogen receptor modulators include, but are not limited to, estrogen, progestogen, estradiol, droloxifene, raloxifene, lasofoxifene, TSE-424, tamoxifen, idoxifene, LY353381, LY117081, toremifene, fulvestrant, 4-[7-(2,2- dimethyl-l-oxopropoxy-4-methyl-2-[4-[2-(l-piperidinyl)ethoxy]phenyl]-2H-l-benzopyran-3-yl]-phenyl- 2,2-dimethylpropanoate, 4,4'-dihydroxybenzophenone-2,4-dinitrophenyl-hydrazone, and SH646.
  • estrogen receptor beta modulator is a compound that selectively agonizes or antagonizes estrogen receptor beta (ERD Agonizing ERD increases transcription of the tryptophan hydroxylase gene (TPH, the key enzyme in serotonin synthesis) via an ERD mediated event.
  • ERD Agonizing ERD increases transcription of the tryptophan hydroxylase gene (TPH, the key enzyme in serotonin synthesis) via an ERD mediated event.
  • TPH tryptophan hydroxylase gene
  • Estrogen receptor modulators refers to compounds which interfere or inhibit the binding of androgens to the receptor, regardless of mechanism.
  • Examples of androgen receptor modulators include finasteride and other 5oc-reductase inhibitors, nilutamide, flutamide, bicalutamide, liarozole, and abiraterone acetate.
  • An inhibitor of osteoclast proton ATPase refers to an inhibitor of the proton ATPase, which is found on the apical membrane of the osteoclast, and has been reported to play a significant role in the bone resorption process. This proton pump represents an attractive target for the design of inhibitors of bone resorption which are potentially useful for the treatment and prevention of osteoporosis and related metabolic diseases. See C.
  • HMG-CoA reductase inhibitors refers to inhibitors of 3-hydroxy-3-methylglutaryl- CoA reductase.
  • Compounds which have inhibitory activity for HMG-CoA reductase can be readily identified by using assays well-known in the art. For example, see the assays described or cited in U.S. Patent 4,231,938 at col. 6, and WO 84/02131 at pp. 30-33.
  • HMG-CoA reductase inhibitor and “inhibitor of HMG-CoA reductase” have the same meaning when used herein.
  • HMG-CoA reductase inhibitors include but are not limited to lovastatin (MEVACOR®; see U.S. Patent Nos. 4,231,938, 4,294,926 and 4,319,039), simvastatin (ZOCOR®; see U.S. Patent Nos. 4,444,784, 4,820,850 and 4,916,239), pravastatin (PRAVACHOL®; see U.S. Patent Nos.
  • HMG-CoA reductase inhibitor as used herein includes all pharmaceutically acceptable lactone and open-acid forms (i.e., where the lactone ring is opened to form the free acid) as well as salt and ester forms of compounds which have HMG-CoA reductase inhibitory activity, and therefor the use of such salts, esters, open-acid and lactone forms is included within the scope of this invention.
  • An illustration of the lactone portion and its corresponding open-acid form is shown below as structures I and ⁇ .
  • HMG-CoA reductase inhibitors where an open-acid form can exist, salt and ester forms may preferably be formed from the open-acid, and all such forms are included within the meaning of the term "HMG-CoA reductase inhibitor" as used herein.
  • the HMG-CoA reductase inhibitor is selected from lovastatin and simvastatin, and most preferably simvastatin.
  • the term "pharmaceutically acceptable salts" with respect to the HMG-CoA reductase inhibitor shall mean nontoxic salts of the compounds employed in this invention which are generally prepared by reacting the free acid with a suitable organic or inorganic base, particularly those formed from cations such as sodium, potassium, aluminum, calcium, lithium, magnesium, zinc and tetramethylammonium, as well as those salts formed from amines such as ammonia, ethylenediamine, N-methylglucamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N- benzylphenethylamine, 1 -p-chlorobenzyl-2-pyrrolidine- 1 ' -yl-methylbenz-imidazole, diethylamine, piperazine, and tris(hydroxymethyl) aminomethane.
  • a suitable organic or inorganic base particularly those formed from
  • salt forms of HMG-CoA reductase inhibitors may include, but are not limited to, acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynapthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamao
  • Ester derivatives of the described HMG-CoA reductase inhibitor compounds may act as prodrugs which, when absorbed into the bloodstream of a warm-blooded animal, may cleave in such a manner as to release the drug form and permit the drug to afford improved therapeutic efficacy.
  • integrin receptor antagonists refers to compounds which selectively antagonize, inhibit or counteract binding of a physiological ligand to the ⁇ v ⁇ 3 integrin, to compounds which selectively antagonize, inhibit or counteract binding of a physiological ligand to the ⁇ v ⁇ 5 integrin, to compounds which antagonize, inhibit or counteract binding of a physiological ligand to both the ⁇ v ⁇ 3 integrin and the ⁇ v ⁇ 5 integrin, and to compounds which antagonize, inhibit or counteract the activity of the particular integrin(s) expressed on capillary endothelial cells.
  • the term also refers to antagonists of the ⁇ v ⁇ 6 > ⁇ *v ⁇ 8 > ⁇ l ⁇ l > ⁇ 2 ⁇ l> ⁇ 5 ⁇ l> ⁇ 6 ⁇ l and ⁇ 4 integrins.
  • the term also refers to antagonists of any combination of ⁇ v ⁇ 3, ⁇ v ⁇ s , ⁇ v ⁇ 6> ⁇ *v ⁇ 8 > ⁇ l ⁇ l > ⁇ 2 ⁇ l > ° ⁇ 5 ⁇ l> ⁇ 6 ⁇ l and ⁇ 6 ⁇ 4 integrins. H.N.
  • the o and ⁇ integrin subunits interact non-covalently and bind extracellular matrix ligands in a divalent cation-dependent manner.
  • the most abundant integrin on osteoclasts is v ⁇ 3 (>10 7 /osteoclast), which appears to play a rate-limiting role in cytoskeletal organization important for cell migration and polarization.
  • the ⁇ v ⁇ 3 antagonizing effect is selected from inhibition of bone resorption, inhibition of restenosis, inhibition of macular degeneration, inhibition of arthritis, and inhibition of cancer and metastatic growth.
  • An osteoblast anabolic agent refers to agents that build bone, such as PTH.
  • parathyroid hormone or its amino-terminal fragments and analogues have been shown to prevent, arrest, partially reverse bone loss and stimulate bone formation in animals and humans.
  • PTH parathyroid hormone
  • Studies have demonstrated the clinical benefits of parathyroid hormone in stimulating bone formation and thereby increasing bone mass and strength. Results were reported by RM Neer et al., New Eng JMed 344 1434-1441, 2001.
  • parathyroid hormone-related protein fragments or analogues, such as PTHrP- (1-36) have demonstrated potent anticalciuric effects [see M.A.
  • administration means introducing the compound or a prodrug of the compound into the system of the animal in need of treatment.
  • a compound of the invention or prodrug thereof is provided in combination with one or more other active agents (e.g., a cytotoxic agent, etc.)
  • administration and its variants are each understood to include concurrent and sequential introduction of the compound or prodrug thereof and other agents.
  • the present invention includes within its scope prodrugs of the compounds of this invention. In general, such prodrugs will be functional derivatives of the compounds of this invention which are readily convertible in vivo into the required compound.
  • the term "administering” shall encompass the treatment of the various conditions described with the compound specifically disclosed or with a compound which may not be specifically disclosed, but which converts to the specified compound in vivo after administration to the patient.
  • Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs,” ed. H. Bundgaard, Elsevier, 1985, which is incorporated by reference herein in its entirety. Metabolites of these compounds include active species produced upon introduction of compounds of this invention into the biological milieu.
  • composition is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.
  • therapeutically effective amount means that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician.
  • treating or “treatment” of a disease as used herein includes: preventing the disease, i.e.
  • the term "bone resorption,” as used herein, refers to the process by which osteoclasts degrade bone.
  • the present invention also encompasses a pharmaceutical composition useful in the treatment of osteoporosis or other bone disorders, comprising the administration of a therapeuticaily effective amount of the compounds of this invention, with or without pharmaceutically acceptable carriers or diluents.
  • compositions of this invention include aqueous solutions comprising compounds of this invention and pharmacologically acceptable carriers, e.g., saline, at a pH level, e.g., 7.4.
  • pharmacologically acceptable carriers e.g., saline
  • the solutions may be introduced into a patient's bloodstream by local bolus injection.
  • the daily dosage will normally be determined by the prescribing physician with the dosage generally varying according to the age, weight, and response of the individual patient, as well as the severity of the patient's symptoms.
  • a suitable amount of compound is administered to a mammal undergoing treatment for a cathepsin dependent condition.
  • Oral dosages of the present invention when used for the indicated effects, will range between about 0.01 mg per kg of body weight per day (mg/kg/day) to about 100 mg/kg/day, preferably 0.01 to 10 mg/kg/day, and most preferably 0.1 to 5.0 mg/kg/day.
  • the compositions are preferably provided in the form of tablets containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100 and 500 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated.
  • a medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably, from about 1 mg to about 100 mg of active ingredient.
  • the most preferred doses will range from about 0.1 to about 10 mg/kg/minute during a constant rate infusion.
  • compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily.
  • preferred compounds for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in the art.
  • the dosage administration will, of course, be continuous rather than intermittant throughout the dosage regimen.
  • the compounds of the present invention can be used in combination with other agents useful for treating cathepsin-mediated conditions.
  • the scope of the invetion therefore encompasses the use of the instantly claimed compounds in combination with a second agent selected from: an organic bisphosphonate; an estrogen receptor modulator; an androgen receptor modulator; an inhibitor of osteoclast proton ATPase; an inhibitor of HMG-CoA reductase; an integrin receptor antagonist; an osteoblast anabolic agent, such as PTH; and the pharmaceutically acceptable salts and mixtures thereof.
  • a second agent selected from: an organic bisphosphonate; an estrogen receptor modulator; an androgen receptor modulator; an inhibitor of osteoclast proton ATPase; an inhibitor of HMG-CoA reductase; an integrin receptor antagonist; an osteoblast anabolic agent, such as PTH; and the pharmaceutically acceptable salts and mixtures thereof.
  • a palladium-catalyzed Suzuki coupling with an appropriate boronic acid provides additional compounds of the current invention.
  • a copper-catalyzed or palladium-catalyzed Buchwald coupling with a suitable amine provides additional compounds of the current invention.
  • a palladium-catalyzed caboxylation followed by amide formation with a suitable amine provides additional compounds of the current invention.
  • a ketone or aldehyde may be condensed with an amino alcohol to give a cyclic aminal. Treatment with 3 equivalents of a Grignard reagent or organolithium reagent will provide the appropriate alkylated amino alcohol. Oxidation of the alcohol with a chromium system such as a Jones oxidation or H 5 K Cr0 3 , or alternatively by a two-step oxidation (eg oxalyl chloride/ DMSO/Et 3 N followed by NaClO) will provide the corresponding carboxylic acid. Peptide coupling and Suzuki reaction as described in Scheme 1 will provide compounds of the current invention. SCHEME 2
  • Compounds of the current invention may also be prepared according to Scheme 4.
  • An appropriately substituted acetate may be enolized with a suitable base (including, but not limited to LDA, KHMDS, NaH or nBuLi) and treated with paraformaldehyde to generate the diol.
  • This diol may be converted to the difluoride using a fluorinating reagent such as DAST. Hydrolysis of the ester followed by Curtius rearrangement will then provide the amine.
  • This amine can displace an appropriately substituted alpha- bromo ester to provide the alpha-amino ester. This may be converted into compounds of the current invention by the method described in Scheme 1.
  • the 4-fluoroleucinol can also be synthesized according to Scheme 8. 4,5-Dehydroleucine is converted to (45)-4-(2-methylprop-2-enyl)-l,3-oxazolidin-2-one as described in the scheme below. This intermediate is then treated with a hydrofluorination reagent such as HF-pyridine to give (45)-4-(2-fluoro-2- methylpropyl)-l,3-oxazolidin-2-one. Basic hydrolysis (i.e. Ba(OH) 2 or NaOH) then affords (2S)-2- amino-4-fluoro-4-methylpentan- 1 -ol.
  • a hydrofluorination reagent such as HF-pyridine
  • the amino alcohols used for the present invention can also be synthesized according to Scheme 10.
  • a protected amino acid is reduced with a reducing agent such as NaBE , with or without an additive such as LiCl, in a solvent such as EtOH or a mixed solvent system such as EtOH and THF.
  • the amino protecting group is then removed with the appropriate method according to the nature of the protecting group.
  • the isolated 5,5,5-trifluoro-L-leucine is then protected with a protecting group such as benzyl carbamate and the carboxylic acid group is esterified.
  • a protecting group such as benzyl carbamate
  • carboxylic acid group is esterified.
  • the two diastereomers at the 4-position are then separated by flash column chromatography.
  • One of the enantiomers, the (2S,4S) protected amino acid is then converted to the amino alcohol as described in scheme 10.
  • the alkylated aminoalcohol is then converted into compounds of the current invention either by the method described in Scheme 2 or by first conducting the Suzuki reaction with the boronic ester of the formula R 7 -B(OH) 2 , then oxidizing the alcohol with a suitable oxidizing agent such as H 5 I ⁇ 6 /Cr0 3 to give the acid and finally treating the acid with an aminoacetonitrile under peptide coupling conditions as described previously.
  • a suitable oxidizing agent such as H 5 I ⁇ 6 /Cr0 3
  • Acids shown in Schemes 1, 2, 6 and 15 may also be prepared as shown in Scheme 16.
  • An appropriately substituted benzyl bromide, iodide or triflate (which may be chiral or racemic) may be coupled with an alpha amino ester under basic conditions. Hydrolysis with aqueous base then provides the acid which can be converted into examples of the current invention.
  • Step 1 Methyl N- ⁇ ('4-bromophenyl)r4-(methylsulfonyl)phenyllmethylenel-L-leucinate
  • 4-bromophenyl)[4-(methylsulfonyl)phenyl]methanone 202 mg, 0.59 mmol
  • L-leucine methyl ester hydrochloride 328 mg, 2.0 mmol
  • camphor sulfonic acid 52 mg, 0.22 mmol
  • Step 2 Methyl N-l (4-bromophenyl)r4-(methylsulfonyl)phenvnmethyl ⁇ -L-leucinate
  • a 1 : 1 mixture of methyl N- ⁇ (4-bromophenyl)[4- (methylsulfonyl)phenyl]methylene ⁇ leucinate and (4-bromophenyl)[4-(methylsulfonyl)phenyl]methanone from step 1 (185 mg, -0.2 mmol) in acetic acid/methanol (1:3, 4 mL) was added sodium borohydride (-400 mg) by portions every 30 min over 2 days (addition was stopped during the night) using a solid addition funnel.
  • Step 3 N- ⁇ (4-bromophenvDr4-(methylsulfonyl)phenyllmethyll-L-leucine
  • methyl N- ⁇ (4-bromophenyl)[4-(methylsulfonyl)phenyl]methyl ⁇ -L- leucinate from step 2 (81 mg, 0.17 mmol) in THF (1 mL) and MeOH (0.5 mL) was added IN LiOH (0.3 mL, 0.3 mmol).
  • the resulting mixture was stirred at room temperature for 18 hours and then partitioned between EtOAc and water + IN HCI (0.5 mL).
  • the organic layer was dried over ⁇ a 2 S0 4 , filtered and concentrated in vacuo to give the title compound as a colorless gum.
  • Step 4 ⁇ - ⁇ -bromophenyD ⁇ -fmethylsulfony phenynmethyll-N ⁇ cvanomethvD-L- leucinamide
  • N- ⁇ (4-bromophenyl)[4-(methylsulfonyl)phenyl]methyl ⁇ -L-leucine from step 3 76 mg, 0.17 mmol
  • HATU 146 mg, 0.38 mmol
  • aminoacetonitrile hydrochloride 52 mg, 0.56 mmol
  • DMF 1.1 mL
  • NN-diisopropylethylamine 0.13 mL, 0.75 mmol
  • Step 5 N J -(cvanomethyl)-N 2 - ⁇ r4-(methylsulfonyl ' )phenylll4'-( ' methylthio)-l.r-biphenyl-4- yllmethyl ⁇ -L-leucinamide
  • N 2 - ⁇ (4-bromophenyl)[4-(methylsulfonyl)phenyl]methyl ⁇ -N i - (cyanomethyl)-L-leucinamide from step 4 72 mg, 0.15 mmol
  • 4-(methylthio)phenylboronic acid 37 mg, 0.22 mmol
  • Step 6 N i -(CvanomethylVN 2 -ir4'-(methylsulfonylVl.l'-biphenyl-4-ylll4- (methylsulfonyl)phenyllmethvU-L-Leucinamide
  • N 7 a solution of N 7 -(cyanomethyl)-N 2 - ⁇ [4-(methylsulfonyl)phenyl][4'-(methylthio)-l,r- biphenyl-4-yl]methyl ⁇ -L-leucinamide (63 mg, 0.12 mmol), sodium tungstate dihydrate (2 mg, 0.006 mmol), tetrabutylammonium hydrogensulfate (4 mg, 0.01 mmol) was added a solution of 30% w/v aqueous hydrogen peroxide (100 ⁇ L, 0.9 mmol) and the resulting mixture was stirred at room temperature for 10 min.
  • Step 1 (2S)-l- ⁇ rfert-butyl(dimethyl silyl1oxyl-4-methylpentan-2-amine
  • L-leucinol 6.0 g
  • DMAP 0.1 g
  • t-butyldimethylsilyl chloride 8.5 g
  • the mixture was stirred at room temperature for 2 hours and then water was added.
  • the organic layer was separated and the aqueous further extracted with dichloromethane.
  • the combined organic layers were washed with brine, dried with magnesium sulfate and the solvent was removed in vacuo to yield the title compound, a residue which was used as such in the next reaction.
  • Step 2 (2y)-l- ⁇ rtert-butyl( ' dimethvDsilylloxyl-4-methyl-N-r ⁇ £ , )-2.2.2- trifluoroethylidenelpentan-2-amine
  • a toluene (300 mL) solution of (25)-l- ⁇ [tert-butyl(dimethyl)silyl]oxy ⁇ -4-methylpentan- 2-amine from Step 1 (50 g) and tifluoroacetaldehyde methyl hemiacetal ( 35 mL) was heated to reflux for 16 hours during which time water was collected in a Dean-Stark trap. The solvent was evaporated in vacuum and the residue was purified on Si0 2 using hexanes and ethyl acetate (9:1) as eluant to yield the title compound.
  • Step 3 (2SV2-(r ⁇ SVl-f4-bromophenylV2.2.2-trifluoroethyllarninol-4-methylpentan-l-ol n-BuLi (2.5 M in hexanes, 42 mL) was added to a -70 °C THF (400 mL) solution of 1,4- dibromobenzene (25.8 g) and the mixture was stirred for 25 minutes.
  • Step 4 (2SV4-methyl-2- ⁇ SV2.2.2-trifluoro-l-r4'-fmethylthio)-l.l'-biphenyl-4- yl1ethyl)amino ' )pentan-l-ol
  • a stream of nitrogen was passed through a suspension made of the bromide from Step 3 (27.7 g), 4-(methylthio)phenylboronic acid (15.7 g), 2 M Na 2 C0 3 (100 mL) and n-propanol (500 mL) for 15 minutes.
  • a 1:3 mixture (3.5 g) of Pd(OAc) 2 and PPh 3 was then added and the reaction was warmed to 70 °C and stirred under nitrogen for 8 hours.
  • Step 5 (2SV4-methyl-2-(((lS 2.2.2-trifluoro-l-14'-( " methylsulfonylVl.r-biphenyl-4- yll ethyl I amino)pentan- 1 -ol
  • Na 2 W0 4 »2H 2 0 (0.16 g) and Bu 4 NHS0 4 (0.81 g).
  • 30 % hydrogen peroxide (12.2 mL) was slowly added and the mixture was stirred at room temperature for 4.5 hours.
  • the mixture was poured slowly on a mixture of ice, dilute aqueous sodium thiosulfate and ethyl acetate.
  • the organic layer was separated and the aqueous further extracted with ethyl acetate (2 X 100 mL).
  • the combined organic layers were washed with brine, dried with magnesium sulfate and the solvent were removed in vacuo to yield a residue which was purified purified on Si0 2 using ethyl acetate and hexanes (1:1) as eluant to yield the product.
  • Step 6 Preparation of N-i ⁇ SV2.2.2-trifluoro-l-r4'-( " methylsulfonylVl.l'-biphenyl-4-vnethyll- L-leucine
  • a suspension of H 5 I0 6 /Cr0 3 (529 mL of 0.44 M in CH3CN; see Note below) was cooled to 0 °C and a solution of the alcohol from Step 5 (20 g) in CH 3 CN (230 mL) was added dropwise. The mixture was stirred at 0-5 °C for 3.5 hours. It was poured into pH 4 Na 2 HP0 4 (1.5 L) under vigorous stirring and the mixture was extracted with diethyl ether (3 X 250 mL).
  • the combined ether extracts were washed with water and brine (1:1), with dilute aqueous NaHS0 3 and brine.
  • the organic extract was dried with sodium sulfate, filtered and the solvents were evaporated to dryness to yield a residue that was split into two batches for the following purification.
  • the crude acid from above (10 g) was dissolved in isopropyl acetate (250 mL) and extracted into cold 0.1 N NaOH (3 X 250 mL).
  • the combined extracts were washed with diethyl ether (250 mL) and then slowly acidified with 6 N HCI to pH 4.
  • the carboxylic acid was extracted with isopropyl acetate (2 X 250 mL) and the isopropyl acetate layer dried and concentrated to yield the product essentially pure and used as such in the next step.
  • the oxidizing reagent HsIO ⁇ /Cr0 3
  • HPLC grade CH 3 CN contains 0.5% water
  • Step 7 Preparation of NVcvanomethvn- ⁇ KlS ⁇ -trifluoro-l- ⁇ '-rmethylsulfonylVl.l'- biphenyl-4-yllethyl 1 -L-leucinamide
  • a DMF (200 mL) solution of the acid from Step 7 (9 g) was added benzotriazol-1- yloxytris(dimethylamino)phosphoniumhexafluorophosphate (11.6 g), aminoacetonitrile hydrochloride (3.94 g) and the mixture was cooled to 0 °C.
  • Triethylamine (9.9 mL) was added dropwise and the mixture warmed to room temperature and stirred for 16 hours.
  • Step 1 Benzyl (3S)-3-r( " tert-butoxycarbonyl)amino1-4-hvdroxybutanoate ⁇ -(tert-Butoxycarbonyl)-L-aspartic acid 4-benzyl ester (30 g) was dissolved in dimethoxyethane (90 mL) and the solution was cooled to -5 °C. N-Methylmorpholine (10.32 mL) was added followed by isobutyl chloroformate (12.7 mL) in such a way to keep the temperature below -10 °C. The mixture was aged for 0.5 hour. The solids were quickly filtered and washed with dimethoxyethane (90 mL).
  • Step 2 Benzyl r(4 ⁇ $ r )-2-oxo- 3-oxazolidin-4-ynacetate
  • dichloroethane 925 mL
  • pyridine 625 mL
  • Anhydrous p-toluenesulfonic anhydride 105.7 g. was added and the mixture was warmed to room temperature and stirred for 1 hour. It was then heated to 90 °C for 2 hours.
  • the mixture was cooled, diluted with dichloromethane (1000 mL) and washed with IN HCI (3 X 600 mL).
  • Step 3 (4S)-4-(2-Hvdroxy-2-methylpropyl " )-1.3-oxazolidin-2-one.
  • Methylmagnesium bromide (227 mL of 3M solution in diethyl ether) was added to a mixture of toluene (340 mL) and THF (340 mL) at -20 °C.
  • the ester from Step 2 (40 g) as a warm THF solution (170 mL) was then added dropwise maintaining the temperature below -10 °C and the mixture was aged for 2 hours.
  • the mixture was then slowly added to a mixture of water (1000 mL) and acetic acid (200 mL) and the mixture was stirred for 2 hours at room temperature.
  • the aqueous layer was separated and the organic extracted with water (2 X 200 mL).
  • the product was extracted from the combined aqueous layers using dichloromethane and a continuous extractor.
  • the dichloromethane extract was evaporated to dryness with the help of heptane.
  • the residue was purified by chromatography on Si0 2 using ethanol and dichloromethane (1:30) to yield (45)-4-(2-hydroxy-2-methylpropyl)-l,3- oxazolidin-2-one.
  • Step 4 (4S)-4-(2-Fluoro-2-methylpropyl)-1.3-oxazolidin-2-one.
  • the alcohol (47.8 g.) from Step 3 as a dichloromethane (100 mL) solution was added to a -70 °C solution of (diethylamino)sulfur trifluoride (48.5 g.) in dichloromethane (500 mL).
  • the mixture was warmed to room temperature and stirred for 1 hour. It was then carefully added to a 0 °C mixture of saturated aqueous NaHC0 3 (800 mL).
  • the organic layer was separated and washed with saturated aqueous NaHC0 3 .
  • Step 5 (2S)-2-Amino-4-fluoro-4-methylpentan-l-ol.
  • fluoro derivative (21.0 g) from Step 4 dissolved in 90% aqueous ethyl alcohol (216 mL) was added potassium hydroxide (21.9 g). The mixture was heated at reflux for 4 hours and cooled to room temperature. The mixture was then concentrated and co-evaporated with toluene (3 X 300 mL). The residue was dissolved in dichloromethane (500 mL) and stirred for 0.5 hour. The suspension was filtered through celite and the celite was washed with dichloromethane (3 X 100 mL).
  • Step 6 (2S)- 1 - ⁇ rtert-butyKdimethyDsilyH oxy ⁇ -4-fluoro-4-methylpentan-2-amine
  • the amino alcohol (21.0 g) from Step 5 was dissolved in dichloromethane (300 mL) and the solution was cooled to 0 °C.
  • 4-(Dimethylamino)pyridine (0.051 g.) and tert-butyldimethylsilyl chloride (21 g.) were added followed by triethylamine (25 mL). The mixture was stirred at room temperature overnight. The reaction mixture was slowly poured into 0 °C saturated aqueous ammonium chloride and extracted with dichloromethane (3 X 300 mL).
  • Step 7 (2SVl-irtgrt-butyl( ' dimethyl silylloxyl-4-fluoro-4-methyl-N-r(lE)-2.2.2- trifluoroethylidenelpentan-2-amine.
  • benzene 126 mL
  • trifluoroacetaldehyde methyl hemiacetal 21.6 mL.
  • the solution was heated at reflux overnight using a Dean-Stark trap to collect water.
  • the reaction mixture was cooled to room temperature and concentrated to dryness.
  • Step 8 ( • 2y)-2-irdS -l-(4-BromophenvD-2.2.2-trifluoroethvnaminol-4-fluoro-4-methylpentan- l-ol.
  • 1,4-dibromobenzene (0.26 g) in THF (4 mL)
  • n-BuLi (0.42 mL of a 2.5M hexanes solution
  • the imine (0.329 g.) from Step 7 in THF (2 mL) was added and the mixture was aged 2 hours.
  • n-Tetrabutylammonium fluroride (6 mL from a IM THF solution) was added and the mixture was stirred at + 5 °C for 16 hrs. It was poured into a mixture of water (50 mL), ammonium chloride (1 g.) and crushed ice and the organic layer was separated. The aqueous was further extracted with ethyl acetate ( 2X 15 mL) and the combined organic layers were dried and concentrated.
  • Step 9 N-rdSVl-(4-bromophenyl)-2.2.2-trifluoroethyll-4-fluoro-L-leucine.
  • a suspension of H 5 I0 6 /Cr0 3 (66 mL of 0.44 M in CH 3 C ⁇ ; Note) was cooled to 0 °C and a solution of the alcohol from Step 8 (1.55 g) in CH 3 CN (5 mL) was added dropwise. The mixture was stirred at 0-5 °C for 3.5 hours. It was poured into pH 4 Na 2 HP0 (200 mL) under vigorous stirring and the mixture was extracted with diethyl ether (3 X 50 mL).
  • the oxidizing reagent (H 5 I0 6 /Cr0 3 ) was prepared as described in Tetrahedron Letters 39 (1998) 5323-5326 but using HPLC grade CH 3 C ⁇ (contains 0.5% water); no water was added.
  • Step 10 N ⁇ r ⁇ yt-l- ⁇ -bromophenylV ⁇ . ⁇ -trifluoroethyll-N'-d-cyanocvclopropyD ⁇ -fluoro-L- leucinamide.
  • Diisopropylethylamine (4.2 mL) was added to a 0 °C suspension of the acid (1.5 g) from Step 9, 1-amino-l-cyclopropanecarbonitrile hydrochloride (1.18 g), 0-(7-azabenzotriazol-l-yl)-N, N, N', N'-tetramethyluroniumhexafluorophosphate (1.94 g) and dimethylformamide (5 mL) and the mixture was reacted at room temperature for 48 hrs. It was then poured on ice and dilute aqueous ammonium chloride.
  • Step 11 N 1 -d-cvanocvclopropylV4-fluoro-N 2 -(dS)-2.2.2-trifluoro-l-r4'-('methylthio)-l.l'- biphenyl-4-yll ethyl ⁇ -L-leucinamide.
  • a stream of nitrogen was passed through a suspension made of the bromide from Step 10 (0.338 g.), 4-(methylthio)phenylboronic acid (0.252 g), 2M ⁇ a 2 C0 3 (0.8 mL) and DMF (4 mL) for 15 minutes.
  • PdCl 2 • dppf (0.1 g) was then added and the reaction was warmed to 85 °C and stirred under nitrogen for 5 hours.
  • the mixture was cooled to room temperature, diluted with ethyl acetate (10 mL) and poured into water (50 mL) and ice. The ethyl acetate layer was separated and the aqueous further extracted with ethyl acetate. The combined ethyl acetate extracts were dried and the solvents removed in vacuo.
  • Step 12 Preparation of N'-d -cvanocvclopropyD-4-fluoro-N 2 -(dS 2.2.2-trifluoro-l-r4'- (methylsulfonvD-l . -biphenyl-4-yllethyl I -L-leucinamide
  • a 0 ° solution of the sulfide (0.265 g) from Step 11 in toluene (5 mL) and dichloromethane (5 mL) was added ⁇ a 2 W0 4 »2H 2 0 (0.002 g) and n-Bu 4 NHS0 4 (0.01 g).
  • Step 1 N 2 -d-
  • ⁇ 2 -[l-(4-bromophenyl)-2,2,2-trifluoroethyl]- ⁇ l-(cyanomethyl)-L- leucinamide (example 2) (100 mg, 0.25 mmol), tris(dibenzylideneacetone)dipalladium (2.3mg, 0.0025mmol), biphenyl-2-yl(di-tert-butyl)phosphine (3mg, 0.01 mmol), potassium phosphate (74 mg, 0.35 mmol) and tert-butyl piperazine-1-carboxylate (56 mg, 0.3 mmol) in dimethoxyethane (0.5 mL) was cooled to
  • Step 2 N ⁇ CyanomethvD-N ⁇ -trifluoro- 1 -( " 4-piperazin- 1 -ylphenyDethvil -L-leucinamide.
  • methanesulfonic acid 53 uL, 0.82mmol
  • Ethyl acetate was added, then the mixture was washed with saturated sodium bicarbonate, brine, dried over magnesium sulfate, filtered and the solvent evaporated under vacuum. Purification by silica gel chromatography eluting with 93% dichloromethane, 0.6% ammonium hydroxide and 6.4% methanol afforded the title compound.
  • Step 1 Preparation of l-(4-bromophenyl)cyclopropanecarbonitrile To a room temperature solution of 4-bromophenylacetonitrile (18.0 g) in 22 mL of sodium hydroxide (50% in water W/W) were added l-bromo-2-chloroethane and (12.0 mL) and benzyltrimethylammonium chloride (627 mg). The mixture was heated at 60 °C overnight. The reaction mixture was cooled to room temperature and diethyl ether was added (300 mL. The ether layer was washed with water (100 mL), hydrogen chloride (100 mL, 10% HCI in water) and brine. The organic layer was dried with magnesium sulfate and the solvent removed in vacuo. The residue was purified by trituration using diethyl ether and hexanes to yield the title compound.
  • Step 2 Preparation of l-(4-bromophenyl)cvclopropanecarboxylic acid
  • a solution of l-(4-bromophenyl)cyclopropanecarbonitrile from Step 1 (13 g) in ethyl alcohol (110 mL) was added a solution of 56 mL of sodium hydroxide (25% ⁇ aOH in water W/W).
  • the mixture was heated at 100 °C overnight. It was cooled to room temperature, poured into ice and hydrogen chloride (1 ⁇ ) and extracted with dichloromethane (2 X 100 mL). The combined extracts were washed with brine, dried with magnesium sulfate and the solvent removed in vacuo to yield the title compound.
  • Step 3 Preparation of l-(4-bromophenv cvclopropanecarboxamide To a -15 °C solution of l-(4-bromophenyl)cyclopropanecarboxylic acid from Step 2 (1.5 g) in chloroform (60 mL) were slowly added isobutyl chloroformate (900 ⁇ L) and triethylamine (1.1 mL). The reaction mixture was stirred at -15 °C for 2 hours.
  • Step 4 Preparation of N 1 -d-cvanocvclopropyl)-4-fluoro-N 2 -(dy)-2.2.2-trifluoro-l-r4-(4.4.5.5- tetramethyl- 1 ,3.2-dioxaborolan-2-yl)phenv ⁇ l ethyl 1 -L-leucinamide
  • a stream of nitrogen was passed through a DMF (40 mL) suspension of N 2 -[(15)-l-(4- bromophenyl)-2,2,2-trifluoroethyl]-N 1 -(l-cyanocyclopropyl)-4-fluoro-L-leucinamide from Example 5, Step 9 (2.0 g), bis(pinacolato)diboron (1.24 g) and potassium acetate (1.53 g) for 15 minutes.
  • the catalyst [1, -bis(diphenylphosphino)-ferrocene]dichloropalladium(IT), complex (1:1) with dichloromethane (181 mg) was then added and the mixture warmed to 65 °C overnight under nitrogen.
  • the mixture was cooled to room temperature, diluted with ethyl acetate and hexanes (1:1, 100 mL) and poured over water (50 mL) and ice (50 g).
  • the organic layer was separated and the aqueous layer further extracted with ethyl acetate and hexanes (1:1, 3 X 50 mL).
  • the combined extracts were washed with brine and dried with magnesium sulfate. Removal of the solvent left a residue which was purified by chromatography on Si0 2 using ethyl acetate and hexanes (1:3 to 1:2) to yield the title compound.
  • Step 5 Preparation of N 2 -((1S)- 1 -( 4-1 " 1 -(aminocarbonyl)cyclopropyllbiphenyl-4-yl I -2,2,2- trifluoroethyl)-N l-cyanocyclopropyl)-4-fluoro-L-leucinamide
  • a stream of nitrogen was passed through a solution of DMF (4 mL) of the boronate from Step 4 (150 mg), l-(4-bromophenyl)cyclopropanecarboxamide from Step 3 (110 mg) and 2 M ⁇ a 2 C0 3 (400 ⁇ L) for 15 minutes.
  • the catalyst [1, r-bis(diphenylphosphino)-ferrocene]dichloropalladium( ⁇ ), complex (1:1) with dichloromethane (12 mg) was then added and the mixture was warmed to 80 °C for 3 hours under nitrogen.
  • the mixture was cooled to room temperature, poured into ice (10 g) and saturated aqueous sodium bicarbonate (20 mL) and extracted with 50 % ethyl acetate (3 X 30 mL). The combined extracts were washed with brine and dried with magnesium sulfate.
  • Step 1 Preparation of methyl N-((benzyloxy)carbonyl)-3-iodo-L-alaninate
  • carbobenzyloxy-L-serine 25 g, 104 mmol
  • ethyl acetate 200 mL
  • diazomethane a solution of diazomethane in ether until a slight yellow color persisted.
  • the solvent was evaporated under vacuum.
  • ⁇ , ⁇ -dimethylformamide 400 mL
  • methyltriphenoxyphosphonium iodide 50 g, 110 mmol
  • Step 2 Preparation of methyl N-((benzyloxy)carbonyl')-4-oxo-L-norvalinate
  • Step 3 Preparation of methyl N-((benzyloxy)carbonyl)-4,4-difluoro-L-norvalinate
  • dichloromethane 20 mL
  • methanol 0.019 mL
  • DAST 2.46 mL
  • the ice bath was removed and replaced with a hot water (57 °C) bath.
  • the hot water bath was replaced 3 times, then the mixture was stirred overnight at room temperature.
  • Step 4 Preparation of benzyl dS)-3,3-difluoro-l-(hychoxymethyl)butylcarbamate
  • benzyl dS 3-,3-difluoro-l-(hychoxymethyl)butylcarbamate
  • Step 5 Preparation of (2S)-l-((fert-butyl(dimethyl)silyl)oxy)-4.4-difluoropentan-2-amine
  • benzyl (15)-3,3-difluoro-l-(hydroxymethyl)butylcarbamate (from Step 4) in ethanol (25 mL) was added palladium on charcoal (10 %, 150 mg) and the mixture was stirred under a H 2 atmosphere (ballon) for 2 h.
  • Dichloromethane was added and the mixture was filtered on celite. The solvent was evaporated under vacuum.
  • Step 6 Preparation of (2 1 y)-l-((tert-butyl(dimethvDsilvDoxyV4.4-difluoro-N-( ' d£ ⁇ -2.2.2- trifluoroethylidene pentan-2-amine
  • (2S)-l-((tert-butyl(dimethyl)silyl)oxy)-4,4-difluoropentan-2-amine, from Step 5 and trifluoroacetaldehyde methyl hemiacetal (80 %, 0.9 mL) in benzene (20 mL) was refluxed over night with a Dean-Stark apparatus.
  • Step 7 Preparation of (2y)-2-((dS)-l- 4-bromophenvD-2.2.2-trifluoroethvnaminoV4.4- difluoropentan- 1 -ol
  • 1,4-dibromobenzene 330 mg
  • THF 5.2 mL
  • 2.5M n-BuLi in hexanes 0.52 mL
  • Step 8 Preparation of N 2 -rdS)-l-(4-bromophenvD-2.2.2-trifluoroethvn-N 1 -d- cyanocyclopropyl)-4,4-difluoro-L-norvalinamide
  • a suspension of H 5 I0 6 /Cr0 3 (27 mL of 0.44 M in CH 3 C ⁇ ; Note) was cooled to 0 °C and a solution of the alcohol from Step 7 (740 mg) in CH 3 CN (10 mL) was added dropwise. The mixture was stirred at 0 °C for 4 hours, with addition of more H s I0 6 /Cr0 3 (2 x 10 mL of 0.44 M in CH 3 CN).
  • Triethylamine (0.42 mL) was added to mixture of the acid (340 mg) from above, 1- amino-1-cyclopropanecarbonitrile hydrochloride (227 mg), benzotriazol-1-yl- oxytripyrrolidinophosphonium hexafluorophosphate (498 mg) and dimethylformamide (4.5 mL) and the mixture was reacted at room temperature for 48 h. It was then poured on dilute sodium bicarbonate. The mixture was extracted with ethyl ether (3x) and the combined organic layers were washed with brine (3x) and dried with magnesium sulfate filtered.
  • Step 1 N 2 -rdSVl-(4-bromophenvn-2.2.2-trifluoroethyll-4-fluoro-N 1 -r( ' 2R.3S)-2-methyl-4- oxotetrahvdrofuran-3-yll-L-leucinamide
  • Triethylamine (0.63 mL) was added to mixture of the acid (500 mg) from Example 5, Step 9, (4S,5R)-4-amino-5-methyldihydrofuran-3(2H)-one hydrochloride (227 mg) (WO 00/69855), benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (744 mg) and dimethylformamide (7 mL) and the mixture was reacted at room temperature for 3 hrs.
  • Step 2 4-fluoro-N 1 -r(2R,35 r )-2-methyl-4-oxotetrahvdrofuran-3-vn-N 2 -fd l y)-2.2.2-trifluoro-l-r4'- (methylthio)biphenyl-4-yri ethyl 1 -L-leucinamide
  • a mixture of the bromide from Step 1 200 mg
  • 4-(methylthio)phenylboronic acid 104 mg
  • 2M aqueous ⁇ a 2 C0 3 (0.51 mL) and DMF (3 mL) for 15 minutes.
  • PdCl 2 dppf 2 (17 mg) was cooled to -78 °C, pumped under high vacuum for 5 minutes, then nitrogen was let into the flask and the mixture was heated at 80 °C and stirred under nitrogen for 3 hours. The mixture was cooled to room temperature, diluted with ethyl acetate washed with saturated ammonium chloride, brine (3x), dried over magnesium sulfate, filtered and the solvent evaporated under vacuum. Purification by silica gel chromatography using 35% ethyl acetate/hexane as eluent afforded the title compound.
  • Step 3 4-fluoro-N 1 -rr2 ?.3y)-2-methyl-4-oxotetrahvdrofuran-3-yll-N 2 -(d 1 y)-2.2.2-trifluoro-l-[4'- (methylsulf ony biphenyl-4-yll ethyl I -L-leucinamide.
  • Step 1 Preparation of 2-amino-2-methylpropanenitrile hydrochloride To a 0 °C solution of ammonium chloride (15.5 g) in water (50 mL) was added a solution of acetone (17 mL) in diethyl ether (50 mL). Then a solution of the sodium cyanide (11.9 g.) in water (35 mL) was slowly added at such a rate that the temperature never exceeds 10 °C. The reaction mixture was stirred for one hour at 0 °C after the addition of the cyanide solution then it was allowed to stand overnight. The ether layer was separated and the aqueous layer was extracted with diethyl ether (2 x 30 mL).
  • Step 2 Preparation of NAl-cvano-l-methylethylVN 2 -! dSy2.2.2-trifluoro-l-r4'- (methylsulf onyl)biphenyl-4-yll ethyl 1 -L-leucinamide
  • Step 6 1.2 g was added benzotriazol-l-yloxytris(dimethylamino)phosphonium hexafluorophosphate (1.55 g), 2-amino-2- methylpropanenitrile hydrochloride from Step 1 (720 mg) and the mixture was cooled to 0 °C.
  • Triethylamine (1.3 mL) was added drop wise and the mixture was warmed to room temperature and stirred for 72 hours. It was poured into ice and saturated aqueous sodium bicarbonate and extracted with diethyl ether (3 X 50 mL). The combined extracts were washed with brine, dried with magnesium sulfate and the solvent removed in vacuo. The residue was purified by chromatography on Si0 2 using a gradient of ethyl acetate and hexanes (1:2 to 1:1) as eluant, followed by trituration using diethyl ether and hexanes to yield the title compound.
  • Step 1 Methyl N-(diphenylmethylene)-4-methylenenorvalinate To a solution of methyl N-(diphenylmethylene)glycinate (12.0 g, 47.4 mmol) in THF (118 mL) at 0 °C was added a solution of IM potassium f ⁇ rt-butoxide in THF (49 mL, 49 mmol). The mixture turned bright yellow and was further stirred for - 15 min. 3-Bromo-2-methylpropene (5.2 mL, 51.3 mmol) was added and the mixture was stirred at room temperature for 2 days. After quenching with water, the mixture was extracted with EtOAc.
  • Step 2 Methyl N-r(benzyloxy)carbonvn-4-methylenenorvalinate A mixture of methyl N-(diphenylmethylene)-4-methylenenorvalinate (6.2 g, 20.2 mmol) from Step 1 and 0.5 M of aqueous HCI (60 mL) was stirred at room temperature overnight. The whole mixture was washed with Et 2 0 (2x). After cooling to 0 °C, IM aqueous ⁇ aOH (40 mL, 40 mmol) was added, followed by EtOAc (50 mL) and benzyl chloroformate (4 mL, 28 mmol). The mixture was stirred at 0 °C for 2h.
  • Step 3 Methyl N-r(benzyloxy)carbonyll-3-d-methylcyclo ⁇ ropyl)alaninate
  • diethylzinc (2 mL, 19.5 mmol)
  • trifluoroacetic acid 1.5 mL, 19.5 mmol
  • a solution of diiodomethane (1.6 mL, 20.0 mmol) in CH 2 C1 2 (8 mL) was added.
  • the mixture was stirred for 15 min and a clear solution resulted.
  • Step 4 Benzyl 2-hvdroxy- 1 -[( 1 -methylcyclopropyDmethyll ethylcarbamate
  • EtOH 40 mL
  • THF 40 mL
  • NaBKU 1.6 g
  • the mixture was stirred at room temperature overnight, quenched with 0.5 M aqueous HCI, extracted with EtOAc.
  • the EtOAc extract was washed with diluted brine (2x), dried (MgS0 4 ) and concentrated. Purification by chromatography gave 2 g of the title compound as a colorless oil.
  • Step 5 2-Amino-3-d-methylcyclopropyl)propan-l-ol
  • the catalyst was filtered off and the filtrate was concentrated to give the title compound.
  • Step 6 N 1 -( ' Cyanomethyl)-3-d-methylcyclopropyl)-N 2 - ⁇ 2.2.2-trifluoro-l-r4'-( ' methylsulfonyl)- 1.1 '-biphenyl-4-yll ethyl ⁇ -alaninamide
  • the title compound was prepared from the aminoalcohol from Step 5 using the same method described in Example 5, Steps 6-12.
  • Step 1 N 2 - 11 -r4-(hvdroxycarbonyl " )phenyll -2,2,2-trifluoroethyl I -N ⁇ fcyanomethvD-L- leucinamide
  • a mixture of dichlorobis(triphenylphosphine)palladium( ⁇ ) (58 mg, 0.08 mmol), triphenylphosphine (155 mg, 0.59 mmol) andN 2 -[l-(4-bromophenyl)-2,2,2-trifluoroethyl]-N 1 - (cyanomethyl)-L-leucinamide Example 2, 1.2 g, 3.0 mmol) in tributylamine (2.5 mL) and water (0.6 mL) was placed in a steel bomb with a teflon coated magnetic bar.
  • the system was purged 3 times with carbon monoxide (100 psi each times) and finally filled with this gas at a pressure of 300 psi.
  • the reaction mixture was heated with continuous stirring at 160°C for 20 hours. Then the system was allowed to cool to room temperature, pressure was released and the resulting residue was partitioned between EtOAc and water + aqueous hydrochloric acid to adjust the pH between 2.5 and 3.0.
  • the organic layer was dried over ⁇ a2S04, filtered and concentrated.
  • the crude product was purified by chromatography using EtOAc, hexane and acetic acid as eluant to give the title compound as a yellow- orange foam.
  • Step 2 N 1 -(cyanomethyl)-N 2 -( ' 2.2,2-trifluoro-l- ⁇ 4-r(4-methylpiperazin-l- yl)carbonyllphenyllethyl)-L-leucinamide
  • N 2 - ⁇ l-[4-(hydroxycarbonyl)phenyl]-2,2,2-trifluoroethyl ⁇ -N 1 - (cyanomethyl)-L-leucinamide from step 1 (480 mg, 1.3 mmol) and benzotriazol-1- yloxytrpyrrolidinophosphonium hexafluorophosphonate (1.35 g, 2.6 mmol) in DMF (8 mL) was slowly added N-methylpiperazine (0.29 mL, 2.6 mmol) followed by triethylamine (0.54 mL, 3.9 mmol).
  • the reaction mixture was stirred at room temperature for 3 hours.
  • the resulting mixture was partitioned between EtOAc and water + aqueous hydrochloric acid to adjust the pH between 2.5 and 3.0.
  • the organic layer was dried over ⁇ a 2 S0 , filtered and concentrated.
  • the crude product was purified by chromatography using MeOH, EtOAc and NH 4 ⁇ H c0nc as eluant to give the title compound as a white foam.
  • Step 1 N ⁇ Ctert-ButoxycarbonyD-N ⁇ methoxy- N'-methyl-L-methioninamide
  • N-Boc-L-methionine 1.0 g, 4.0 mmol
  • 0-(7-azabenzotriazol- l-yl)-N, N, N', N'-tetramethyluronium hexafluorophosphate 3.3 g, 8.7 mmol
  • N,0- dimethylhydroxylamine hydrochloride 1.0 g, 10.2 mmol
  • DMF 15 mL
  • triethylamine 2.2 mL, 15.8 mmol
  • Step 2 tgrt-ButvUdS)-l-r2-(methylthio)ethyll-2-oxopropyllcarbamate
  • a solution of ⁇ -(tert-butoxycarbony ⁇ -N ⁇ methoxy- N ⁇ methyl-L-methioninamide from Step 1 200 mg, 0.68 mmol
  • a solution of methyl lithium 1.4 M in hexane 1.1 mL, 1.5 mmol.
  • the reaction was stirred at this temperature for 2 hours and then cold- quenched with an aqueous solution of ammonium acetate 25% w / v .
  • Step 3 (3S)-3-Amino-5-(methylthio pentan-2-one, hydrochloride
  • a solution hydrogen chloride 4.0 M in 1,4-dioxane (3 mL, 12 mmol).
  • the resulting mixture was stirred at room temperature for 1 hour.
  • the solvent was removed in vacuo and the resulting residue azeotroped with toluene (2 X 10 mL) to give the title compound as a white solid.
  • Step 4 N 1 -(dSVl-r2-rmethylthio ethyll-2-oxopropyll-N 2 -ldSV2.2.2-trifluoro-l-r4'- (methylsulfonyl)biphenyl-4-yll ethyl I -L-leucinamide
  • the reaction mixture was stirred at room temperature for 3 hours.
  • the resulting mixture was partitioned between EtOAc and half-saturated aqueous ⁇ aHC0 3 .
  • the organic layer was dried over Na 2 S0 4 , filtered and concentrated.
  • the crude product was purified by chromatography using EtOAc and hexane as eluant to give the title compound as a white foam.
  • Step 1 Methyl N-(dSV2.2.2-trifluoro-l-r4'-(methylsulfonvDbiphenyl-4-yllethyll-L-leucyl-L- methioninate Using the procedure described for Example 15, Step 4, where (3S)-3-amino-5- (methylthio)pentan-2-one, hydrochloride was substituted for methyl L-methionate hydrochloride, the title compound was obtained and was crystallized from EtOAc and hexane (1:2) to give a white solid. MS (+ESI): 589.3 [M+1] " *
  • Step 2 N-(dSV2,2.2-trifluoro-l-r4'-(methylsulfonvnbiphenyl-4-yllethyll-L-leucyl-L- methionine
  • the reaction mixture was stirred at room temperature for 18 hours.
  • the resulting mixture was partitioned between EtOAc and half-saturated aqueous ⁇ aHC0 3 .
  • the organic layer was dried over Na 2 S0 , filtered and concentrated.
  • the crude product was purified by chromatography using EtOAc and hexane as eluant to give the title compound as a white foam which was crystallized from EtOAc and hexane to give a white solid.
  • Step 4 ( ' 2SV2-(rdSVl-(4-BromophenylV2.2.2-trifluoroethyllaminol-4.4-dichloro-N-d- cyanocyclopropyDbutanamide
  • (2S)-2- ⁇ [(lS)-l-(4-bromophenyl)-2,2,2-trifluoroethyl] amino ⁇ -4,4- dichlorobutanoic acid 275 mg, 0.67 mmol
  • 1 -amino- 1 -cyclopropane carbonitrile hydrochloride 159 mg, 1.34 mmol
  • HATU coupling reagent 305 mg, 0.8 mmol
  • Triethylamine (0.3 mL, 2.1 mmol) was added and the mixture stirred overnight and then poured into NaHC0 3 solution and ethyl acetate. The organic layer was separated, washed with brine, IN HCI and brine again. The organic layer was separated, dried (Na 2 S0 ), filtered and concentrated to give 393 mg of an oil which was purified by column chromatography eluting with 6:3:1 toluene: ethyl acetate: dichloromethane. Swishing the pure product with diethyl ether gave the title compound as a white solid and as a 85:15 mixture of diastereomers.
  • Step 1 Preparation of (2S)- ⁇ - rtert-butylfdimethvnsilylloxy -N-rdZl ⁇ -difluoroethylidenel ⁇ - methylpentan-2-amine
  • 2S)-l- ⁇ [tert-butyl(dimethyl)silyl]oxy ⁇ -4-methylpentan-2-amine (Example 4, Step 1, 8.5 g, 36.8 mmol) and difluoroacetaldehyde ethyl hemiacetal (5.0g, 39.7 mmol) in benzene was refluxed with a Dean-stark trap overnight. Solvent was removed in vacuo. The residue was passed through a short silica column and eluted with hexanes: EtOAc (10: 1) to give the title compound as a pale yellow oil.
  • Step 2 Preparation of (2S)-2- ⁇ rdS)-l-(4-bromophenyl)-2.2-difluoroethyllarnino>-4- methylpentan- 1 -ol 7i-BuLi (2.5 M in hexanes, 1.43 mL) was added to a -70 °C THF (8.5 mL) solution of 1,4-dibromobenzene (884 mg) and the mixture was stirred for 15 minutes.
  • Step 3 Preparation of N-rdS)-l-(4-bromophenyl)-2,2-difluoroethyll-L-leucine
  • a suspension of H 5 I0 6 /Cr0 3 (5.5 mL of 0.40 M in CH3C ⁇ ; see Note below) was cooled to 0 °C and a solution of the alcohol from Step 2 (250 mg) in CH 3 CN (3.7 mL) was added dropwise. The mixture was stirred at 0-5 °C for 3.5 hours. After this period, 2.0 mL of the oxidant were added.
  • the oxidizing reagent (H 5 I0 6 /Cr0 3 ) was prepared as described in Tetrahedron Letters 39 (1998) 5323-5326 but using HPLC grade CH 3 CN (contains 0.5% water); no water was added.
  • HNMR CD 3 COCD 3 ) ⁇ 7.55(2H, d), 7.4(2H, d), 6.05(1H, dt), 3.95-4.05(lH, m), 3.45(1H, t), 2.7- 3.0(broad m, NH70H), 1.85-1.95(1H, m), 1.5(2H, t), 0.95 (3H, d), 0.9(3H, d).
  • Step 4 Preparation of N 2 -rdS)-l-(4-bromophenyl)-2,2-difluoroethvn-N J -d-cvanocvclopropy - L-leucinamide
  • a DMF (2 mL) solution of the acid from Step 3 (258 mg) were added 0-(7- azabenzotriazol-l-yl)- ⁇ , N, N', N'-tetramethyluronium hexafluorophosphate (337 mg), 1- aminocyclopropanecarbonitrile hydrochloride (175 mg).
  • diisopropylethylamine (0.45 mL) was added dropwise and the mixture was stirred for 16 hours.
  • Step 5 Preparation of Nl-d -cvanocvclopropyl)-N2-l(iy)-2.2-difluoro-l-r4-(4.4.5.5-tetramethyl- 1 ,3 ,2-dioxaborolan-2-yl)phenyll ethyl I -L-leucinamide.
  • a DMF (60 mL) solution of the arylbromide from Step 4 (5.23 g) and of bis(pinacolato)diboron (3.8 g) were added potassium acetate (3.7 g) and PdCl 2 dppf (309 mg). A stream of nitrogen was passed through the suspension for 1 minute. The reaction mixture was heated at 80°C for 16h.
  • Step 6 Preparation of N 1 -d-cvanocvclopropylVN 2 - ⁇ (lS ' )-2.2-difluoro-l-r4-(3-methyl-2- thienyDphenyl] ethyl 1 -L-leucinamide.
  • a stream of nitrogen was passed through a suspension made of the aryl boronate from Step 6 (200 mg), 2-bromo-3-methylthiphene (115 mg), 2 M ⁇ a 2 C0 3 (0.65 mL), DMF (4.3 mL) and PdC dppf (11 mg) for 1 minute.
  • the mixture was then heated in microwave (SmithCreator) for 500 seconds (fixed hold time: OFF) at 120°C (absorption level: high). It was cooled to room temperature, diluted with ethyl acetate (20 mL) and poured into a saturated solution of sodium bicarbonate. The ethyl acetate layer was separated and the aqueous further extracted with ethyl acetate (2 X 15 mL). The combined ethyl acetate extracts were washed with brine and dried with magnesium sulfate.
  • Step 1 Benzyl 3-hvdroxy-4-r(N- ⁇ dS)-2,2,2-trifluoro-l-r4'-(methylsulfonyl)biphenyl-4-yl1ethyl
  • ⁇ - ⁇ (lS)-2,2,2-trifluoro-l-[4'-(methylsulfonyl)-l,l'-biphenyl-4- yl] ethyl ⁇ -L-leucine Example 4, Step 6, 605 mg, 1.37 mmol
  • benzyl 4-amino-3-hydroxyazepane-l- carboxylate WO 0134565, J.
  • Step 2 Benzyl 3-oxo-4-r(N-(dS)-2.2.2-trifluoro-l-r4'-(methylsulfonv biphenyl-4-yllethyll-L- leucyDaminol azepane- 1 -carboxylate
  • benzyl 3-hydroxy-4-[(N- ⁇ (lS)-2,2,2-trifluoro-l-[4'- (methylsulfonyl)biphenyl-4-yl] ethyl ⁇ -L-leucyl)amino] azepane- 1 -carboxylate 98 mg, 0.14 mmol
  • dichloromethane 3 mL
  • Step 1 N 1 -(3-hvdroxyazepan-4-ylVN 2 - ⁇ dSV2.2.2-trifluoro-l-r4'-('methylsulfonyl)biphenyl-4- yll ethyl ⁇ -L-leucinamide
  • a mixture of benzyl 3-hydroxy-4-[(N- ⁇ (lS)-2,2,2-trifluoro-l-[4'- (methylsulfonyl)biphenyl-4-yl]ethyl ⁇ -L-leucyl)amino]azepane-l-carboxylate Example 24, Step 1, 710 mg, 1.03 mmol
  • 10% Pd/C (490 mg) in 2: 1 EtOH:EtOAc (80 mL) was flushed with hydrogen and stirred under a hydrogen balloon for 2h.
  • the reaction mixture was filtered through celite and concentrated to give the title compound.
  • Step 2 N 1 -r3-hvdroxy-l-fpyridin-2-ylsulfonyl)azepan-4-yll-N 2 -(dS , )-2.2.2-trifluoro-l-r4'- (methylsulf onyl)biphenyl-4-yll ethyl 1 -L-leucinamide
  • ⁇ -(S-hydroxyaze an ⁇ -y -N ⁇ Kl ⁇ -trifluoro-l- ⁇ '- (methylsulfonyl)biphenyl-4-yl] ethyl ⁇ -L-leucinamide 567 mg, 1.02 mmol
  • dichloromethane 10 mL
  • 2-pyridinesulfonyl chloride 204 mg, 1.15 mmol
  • Step 3 N 1 -r3-oxo-l-(pyridin-2-ylsulfonvDazepan-4-yll-N 2 -ld.y)-2,2.2-trifluoro-l-l4'- (methylsulfonyl)biphenyl-4-yllethyl 1 -L-leucinamide
  • N 1 -[3-hydroxy-l-(pyridin-2-ylsulfonyl)azepan-4-yl]- N 2 - ⁇ (lS)-2,2,2-trifluoro-l-[4'-(methylsulfonyl)biphenyl-4-yl]ethyl ⁇ -L-leucinamide 460 mg, 0.73 mmol
  • dichloromethane 15 mL
  • Dess-Martin periodinane 420 mg, 1.0 mmol
  • Step 1 ( -bromophenyl)(4-methoxyphenyl)methanol
  • THF 80 mL
  • n-butyllithium 16 mL, 2.5M in hexanes
  • p-anisaldehyde 5 g, 37 mmoles in 4.5 mL of THF
  • the reaction mixture was quenched with methanol (5 mL) and saturated aqueous ammonium chloride (100 mL).
  • Step 2 Methyl N-r(4-bromophenylX4-methoxyphenyl)methyl1-L-leucinate (4-Bromophenyl)(4-methoxyphenyl)methanol (1.15 g, 3.9 mmoles) was dissolved in dichloromethane (4 mL) along with tetrabutylammonium bromide (130 mg, 0.4 mmoles).
  • Step 3 N-[('4-bromophenyl)(4-methoxyphenyl)methvn-L-leucine
  • methyl N-[(4-bromopheny ⁇ )(4- methoxyphenyl)methyl]-L-leucinate (1.25g, 3 mmol) in 60 mL of approximately 2:1:1 THF MeOH/Water was added lihium hydroxide monohydrate (250 mg, 6 mmoles). The mixture was stirred overnight and concentrated. The residue was partitioned between dichloromethane (50 mL) and pH 3.5 phosphate buffer (50 mL).
  • Step 4 N 2 -r(4-bromophenyl ⁇ 4-methoxyphenyl)methyn-N 1 -(cyanomethyl)-L-leucinamide
  • N-[(4-bromophenyl)(4-methoxyphenyl)methyl]-L-leucine 149 mg, 0.37 mmol
  • aminoacetonitrile hydrochloride 87 mg, 0.73 mmol
  • HATU 153 mg, 0.403 mmol
  • this determines the molecular fragments corresponding to Ri and R 2 ; 2) for the fragment of the compound corresponding to the formula of the invention, the amine hydrogen forms a hydrogen bond with the oxygen of Gly66, therefore having a distance of less than 4 A between these two atoms; 3)
  • the fragments of the compound corresponding to R 2 and R 3 were placed in the cathepsin subsites S 2 and S 3 , respectively, according to the labeled carbon atoms on the chemical structure for the compound.
  • the carbon labeled "2" in the structure was located in S2 and the carbon labeled "3" was located in S3, such that the distances to cathepsin C ⁇ 's in Table 1 were within an angstrom of those given in Table 1. Although these placements were done manually and approximately, they were placed intentionally to lead to favorable interactions.
  • the enantiomer corresponding to the chemical formula of the invention was used for the calculation. The energy minimization was carried out using the software MacroModel with the MMFFs force field. All atoms of the compound were allowed to move, but for Cathepsin K only protein sidechains having an atom within 6 A of the compound were allowed to move; in this case the whole sidechain could move.
  • Cathepsin K Assay Serial dilutions (1/3) from 500 ⁇ M down to 0.0085 ⁇ M of test compounds were prepared in dimethyl sulfoxide (DMSO). Then 2 ⁇ L of DMSO from each dilution were added to 50 ⁇ L of assay buffer (MES, 50 mM (pH 5.5); EDTA, 2.5 mM; DTT, 2.5 mM and 10% DMSO) and 25 ⁇ L of human cathepsin K (0.4 nM) in assay buffer solution. The assay solutions were mixed for 5-10 seconds on a shaker plate and incubated for 15 minutes at room temperature. Z-Leu-Arg-AMC (8 ⁇ M) in 25 ⁇ L of assay buffer was added to the assay solutions.
  • assay buffer MES, 50 mM (pH 5.5); EDTA, 2.5 mM; DTT, 2.5 mM and 10% DMSO
  • Cathepsin L Assay Serial dilutions (1/3) from 500 ⁇ M down to 0.0085 ⁇ M of test compounds were prepared in dimethyl sulfoxide (DMSO). Then 2 ⁇ L of DMSO from each dilution were added to 50 ⁇ L of assay buffer (MES, 50 mM (pH 5.5); EDTA, 2.5 mM; DTT, 2.5 mM and 10% DMSO) and 25 ⁇ L of human cathepsin L (0.5 nM) in assay buffer solution. The assay solutions were mixed for 5-10 seconds on a shaker plate and incubated for 15 minutes at room temperature. Z-Leu-Arg-AMC (8 ⁇ M) in 25 ⁇ L of assay buffer was added to the assay solutions.
  • assay buffer MES, 50 mM (pH 5.5); EDTA, 2.5 mM; DTT, 2.5 mM and 10% DMSO
  • Cathepsin B Assay Serial dilutions (1/3) from 500 ⁇ M down to 0.0085 ⁇ M of test compounds were prepared in dimethyl sulfoxide (DMSO). Then 2 ⁇ L of DMSO from each dilution were added to 50 ⁇ L of assay buffer (MES, 50 mM (pH 5.5); EDTA, 2.5 mM; DTT, 2.5 mM and 10% DMSO) and 25 ⁇ L of human cathepsin B (4.0 nM) in assay buffer solution. The assay solutions were mixed for 5-10 seconds on a shaker plate and incubated for 15 minutes at room temperature. Z-Leu-Arg-AMC (8 ⁇ M) in 25 ⁇ L of assay buffer was added to the assay solutions.
  • assay buffer MES, 50 mM (pH 5.5); EDTA, 2.5 mM; DTT, 2.5 mM and 10% DMSO

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Abstract

La présente invention a trait à une nouvelle classe de composés de formule (I), dans laquelle R1, R2, R3 and R4 sont tels que définis dans la description, qui sont des inhibiteurs de la cystéine protéase, comprenant mais de manière non exclusive, des inhibiteurs des cathepsines K, L, S et B. Ces composés sont utiles pour le traitement de maladies dans lesquelles l'inhibition de la résorption osseuse est indiquée, telles que l'ostéoporose, l'arthrose et al polyarthrite rhumatoïde.
EP04761741A 2003-08-27 2004-08-23 Inhibiteurs de la cathepsine Withdrawn EP1660436A4 (fr)

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KR20060079143A (ko) 2003-09-18 2006-07-05 액시스 파마슈티컬스 인코포레이티드 시스테인 프로테아제 억제제로서의 할로알킬 함유 화합물
EP1694647B1 (fr) * 2003-12-12 2016-11-09 Merck Canada Inc. Inhibiteurs des protéases à cystéine du type cathepsine
CA2552726A1 (fr) * 2004-01-08 2005-07-21 Merck Frosst Canada Ltd. Inhibiteurs des cysteines proteases de type cathepsine
US20090312277A1 (en) * 2004-11-19 2009-12-17 Abdelhadi Rebbaa Compositions And Methods For Reversing Or Preventing Resistance Of A Cancer Cell To A Cytotoxic Agent
EP1817275A1 (fr) * 2004-12-01 2007-08-15 Schering Aktiengesellschaft Composes contenant un haloalkyle, utilises comme inhibiteurs de cysteine proteases
EP1819667B1 (fr) 2004-12-02 2012-10-17 ViroBay, Inc. Composes de sulfonamide utilises comme inhibiteurs des cysteine proteases
EP1841419A4 (fr) * 2005-01-19 2009-02-25 Merck Frosst Canada Ltd Inhibiteurs de la cathepsine k et obesite
KR101447897B1 (ko) 2005-03-21 2014-10-07 비로베이, 인코포레이티드 시스테인 단백질분해효소 억제제로서의 알파 케토아미드화합물
MX2007011739A (es) 2005-03-22 2008-03-14 Celera Genomics Compuestos que contienen sulfonilo como inhibidores de cisteina proteasa.
WO2007012180A1 (fr) * 2005-07-26 2007-02-01 Merck Frosst Canada Ltd. Inhibiteurs de la cystéine protéase de la famille de la papaïne pour le traitement des maladies parasitaires
EP1937634A4 (fr) * 2005-10-12 2009-08-12 Merck Frosst Canada Ltd Inhibiteur de cysteine proteases de type cathepsine
US20080051380A1 (en) 2006-08-25 2008-02-28 Auerbach Alan H Methods and compositions for treating cancer
ZA200902477B (en) 2006-10-04 2010-08-25 Virobay Inc Di-fluoro containing compounds as cysteine protease inhibitors
US7893112B2 (en) 2006-10-04 2011-02-22 Virobay, Inc. Di-fluoro containing compounds as cysteine protease inhibitors
US8273913B2 (en) 2007-04-02 2012-09-25 Merck Canada Inc. Amidation process for the preparation of cathepsin K inhibitors
BRPI0822420A2 (pt) * 2008-04-01 2014-10-07 Virobay Inc Composto, composição farmaceutica, e, métodos para tratar uma doença em uma animal e para tratar um paciente que passa por uma tarepia.
WO2010142985A1 (fr) * 2009-06-10 2010-12-16 Astrazeneca Ab Composés 761 n-[1-cyano-2-(phényle)éthyle]pipéridine-2-ylcarboxamide substitués
US10130689B2 (en) * 2009-06-22 2018-11-20 Diffusion Pharmaceuticals Llc Diffusion enhancing compounds and their use alone or with thrombolytics
US8324417B2 (en) * 2009-08-19 2012-12-04 Virobay, Inc. Process for the preparation of (S)-2-amino-5-cyclopropyl-4,4-difluoropentanoic acid and alkyl esters and acid salts thereof
WO2012054388A1 (fr) * 2010-10-18 2012-04-26 The Trustees Of Columbia University In The City Of New York Inhibiteurs de tph1 et bisphosphonates destinés à la prévention et au traitement de maladies caractérisées par une faible masse osseuse
CZ2014941A3 (cs) * 2014-12-19 2016-06-29 Zentiva, K.S. Příprava vysoce čistého intermediátu pro syntézu Odanacatibu
CN106866502B (zh) * 2015-12-10 2020-10-09 广东东阳光药业有限公司 组织蛋白酶k抑制剂及其用途
BR112019001398A2 (pt) 2016-07-29 2019-05-07 Janssen Pharmaceutica Nv métodos para tratamento de câncer de próstata
CN110981937B (zh) * 2018-09-30 2021-11-12 北京和理咨询有限公司 Odn或衍生物的多肽偶合物及其制备方法和应用
EP3946332A1 (fr) 2019-04-05 2022-02-09 Université de Bretagne Occidentale Inhibiteurs du récepteur 2 activé par une protéase pour le traitement d'une neuropathie sensorielle induite par une intoxication neurotoxique marine

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CA2535366A1 (fr) 2005-03-10
US20060287402A1 (en) 2006-12-21

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